LOCUS       sequence-208663-        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"
     CDS             357..662
                     /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     541..560
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(740..758)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      754..801
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(869..874,875..891,892..897))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(910..1725)
                     /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
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPVPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1633..1652
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1717..1736
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1847..2435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2336..2355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2512..2531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2512..2530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2562..2584
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2598..2615
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2637..2657
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 actagtaggc ctgtcgacgg tcagtccggc ttactaaaag ccagataaca gtatgcgtat
       61 ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca
      121 aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt
      181 tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga
      241 agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt
      301 cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc
      361 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
      421 gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc
      481 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
      541 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
      601 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
      661 aaatgtcagg ctcccttata cacagccagt cgactgaccg tcaggcctcc atgggagctc
      721 ctgcagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      781 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc ctgactgcgt
      841 tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca
      901 tgagaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
      961 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     1021 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     1081 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     1141 agtgacgact gaatccggtg agaatggcaa aagtttgtgc atttctttcc agacttgttc
     1201 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     1261 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     1321 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     1381 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     1441 ccatgcgtca tcaggagtac ggataaaatg cttgatggtc ggaacaggca taaattccgt
     1501 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     1561 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     1621 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     1681 taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     1741 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt
     1801 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1861 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1921 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1981 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2041 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2101 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2161 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2221 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2281 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2341 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2401 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2461 ggttcctggc cttttgctgg cctttggtac cagatctcga tcccgcgaaa ttaatacgac
     2521 tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa ctttaagaag
     2581 gagatataca tatgggccac catcatcacc accatgatta tgatattcca actaccgaga
     2641 atttgtattt tcagggggga tcc
//
