LOCUS       sequence-172467-        2146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid for ""Tip Snip"" cloning (described in forthcoming 
            Biotechniques article).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2146)
  AUTHORS   Matsumura I
  TITLE     Semi-automated Tip Snip cloning of restriction fragments into and 
            out of plasmid polylinkers.
  JOURNAL   Biotechniques. 2017 Mar 1;62(3):99-106. doi: 10.2144/000114522.
  PUBMED    28298176
REFERENCE   2  (bases 1 to 2146)
  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"; journalName: 
            "Biotechniques"; date: "2017-03-1- 1"; volume: "62"; issue: "3"; 
            pages: "99-106"
COMMENT     Sequence Label: IMBB2.4-pUC57-mini
FEATURES             Location/Qualifiers
     source          1..2146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    109..130
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    150..166
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     155..177
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             201..218
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             225..245
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             291..311
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             318..335
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    345..365
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     rep_origin      complement(464..1052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(544..563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1223..2014,2015..2083))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1846..1865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(2084..2146,1..42))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
       61 gccacctgac gtcccaatga tcaattgggt accttaatta agacgtcgga attcgcggcc
      121 gcttctagag catgcgggcc cgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      181 cagctcatat gggcagcagc catcatcatc atcatcacag atctgaaaac ctgtacttcc
      241 agggtgccat ggccctgagg gccgatatcg gcctatgcgg cctgaagctt gaaaatttat
      301 atttccaggg cggatcccac catcatcacc accattgact cgagtactag tagcggccgc
      361 tgcagcatcg atgtttaaac gagctcatgc atatcggaaa gaacatgtga gcaaaaggcc
      421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
      541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
      601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
      661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
      721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
      841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
      901 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
      961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1081 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     1441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     1501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     1561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     1621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     1681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     1741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     1801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     1861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     1921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     1981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2101 attattgaag catttatcag ggttattgtc tcatgagcgg atacat
//
