LOCUS       sequence-327986-        3832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Two valuable constructs for either IPTG-induced or 
            temperature-induced expression can be simultaneously obtained in a 
            single cloning process by utilizing plasmid pANY4..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3832)
  TITLE     Yingfeng An Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3832)
  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            "Unpublished"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: pANY4
FEATURES             Location/Qualifiers
     source          1..3832
                     /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"
     CDS             complement(879..896)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    974..988
                     /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)"
     RBS             1237..1245
                     /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)"
     CDS             1334..2047
                     /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"
     primer_bind     complement(2061..2079)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2075..2122
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2190..2195,2196..2212,2213..2218))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(2231..3046)
                     /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(2231..3046)
                     /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     2954..2973
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3038..3057
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3168..3756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3657..3676
                     /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 tcctgcaggt gatggtgatg gtgatgcata
      901 tgtatatctc cttaattttt aaccaatgct tcgtttcgta tcacacaccc caaagccttc
      961 tgctttgaat gctgcccttc ttcagggctt aatttttaag agcgtcacct tcatggtggt
     1021 cagtgcgtcc tgctgatgtg ctcagtatca ccgccagtgg tatttatgtc aacaccgcca
     1081 gagataattt atcaccgcag atggttatct gtatgttttt tatatgaatt tattttttgc
     1141 aggggggcat tgtttggtag gtgagagatc tttagctgtc ttggtttgcc caaagcgcat
     1201 tgcataatct ttcagggtta tgcgttgttc catacaacct ccttagtaca tgcaaccatt
     1261 atcaccgcca gaggtaaaat agtcaacacg cacggtgtta gatatttatc ccttgcggtg
     1321 atagatttaa cgtatgagca caaaaaagaa accattaaca caagagcagc ttgaggacgc
     1381 acgtcgcctt aaagcaattt atgaaaaaaa gaaaaatgaa cttggcttat cccaggaatc
     1441 tgtcgcagac aagatgggga tggggcagtc aggcgttggt gctttattta atggcatcaa
     1501 tgcattaaat gcttataacg ccgcattgct tacaaaaatt ctcaaagtta gcgttgaaga
     1561 atttagccct tcaatcgcca gagaaatcta cgagatgtat gaagcggtta gtatgcagcc
     1621 gtcacttaga agtgagtatg agtaccctgt tttttctcat gttcaggcag ggatgttctc
     1681 acctaagctt agaaccttta ccaaaggtga tgcggagaga tgggtaagca caaccaaaaa
     1741 agccagtgat tctgcattct ggcttgaggt tgaaggtaat tccatgaccg caccaacagg
     1801 ctccaagcca agctttcctg acggaatgtt aattctcgtt gaccctgagc aggctgttga
     1861 gccaggtgat ttctgcatag ccagacttgg gggtgatgag tttaccttca agaaactgat
     1921 cagggatagc ggtcaggtgt ttttacaacc actaaaccca cagtacccaa tgatcccatg
     1981 caatgagagt tgttccgttg tggggaaagt tatcgctagt cagtggcctg aagagacgtt
     2041 tggctgattg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     2101 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc cctgactgcg
     2161 ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa gctgtcaaac
     2221 atgagaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     2281 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     2341 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     2401 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     2461 gagtgacgac tgaatccggt gagaatggca aaagtttgtg catttctttc cagacttgtt
     2521 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     2581 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     2641 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     2701 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     2761 accatgcgtc atcaggagta cggataaaat gcttgatggt cggaagcggc ataaattccg
     2821 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     2881 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     2941 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     3001 ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     3061 tactgtttat gtaagcagac agttttattg ttcatgacaa aaatccctta acgtgagttt
     3121 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3181 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3241 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3301 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3361 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3421 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3481 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3541 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3601 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3661 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3721 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3781 cggttcctgg ccttttgctg gcctttggta ccagatctcg atcccgcgaa at
//
