LOCUS       sequence-385594-        2144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid carries a fragment from a 24-fragment lacI/lacZ 
            cassette split that was designed as a Golden Gate assembly with high
            fidelity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2144)
  AUTHORS   Potapov V, Ong JL, Kucera RB, Langhorst BW, Bilotti K, Pryor JM, 
            Cantor EJ, Canton B, Knight TF, Evans TC Jr, Lohman GJS
  TITLE     Comprehensive Profiling of Four Base Overhang Ligation Fidelity by 
            T4 DNA Ligase and Application to DNA Assembly.
  JOURNAL   ACS Synth Biol. 2018 Nov 16;7(11):2665-2674. doi: 
            10.1021/acssynbio.8b00333. Epub 2018 Oct 29.
  PUBMED    30335370
REFERENCE   2  (bases 1 to 2144)
  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: 
            "10.1021/acssynbio.8b00333"; journalName: "ACS Synth Biol"; date: 
            "2018-11-16- 16"; volume: "7"; issue: "11"; pages: "2665-2674"
COMMENT     Sequence Label: pUC57-mini v2-HF24_F15
FEATURES             Location/Qualifiers
     source          1..2144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     89..111
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     103..120
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     104..120
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             148..170
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    185..209
                     /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)."
     promoter        complement(join(213..219,220..237,238..243))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(389..977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(469..488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1148..1939,1940..2008))
                     /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     1771..1790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2009..2113)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccaatgatgg tctcgtcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
       61 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
      121 gaatccgtaa tcatggtcat atgtatatct ccttcttaaa gttaaacaaa attatttcta
      181 gaggggaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      241 aaagcctggg atcgagatct cgatcctcta cgccggacgc atcgtggccg gcatcaccgg
      301 cgccacaggt cgagaccatc ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      361 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      421 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      481 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      541 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      601 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      661 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
      721 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
      781 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
      841 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
      901 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
      961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1021 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1081 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1141 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1201 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1261 ctggccccag tgctgcaatg ataccgcgag atccacgctc accggctcca gatttatcag
     1321 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1381 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     1441 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     1501 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     1561 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     1621 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     1681 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     1741 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     1801 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     1861 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     1921 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     1981 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2041 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2101 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
