LOCUS       sequence-334768-        3430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  test plasmid for 5RM assembly, expresses lacI repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3430)
  AUTHORS   Matsumura I
  TITLE     Golden Gate assembly of BioBrick-compliant parts using Type II 
            restriction endonucleases.
  JOURNAL   Biotechniques. 2022 May;72(5):185-193. doi: 10.2144/btn-2021-0083. 
            Epub 2022 Mar 8.
  PUBMED    35255734
REFERENCE   2  (bases 1 to 3430)
  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: 
            "10.2144/btn-2021-0083"; journalName: "Biotechniques"; date: 
            "2022-05"; volume: "72"; issue: "5"; pages: "185-193"
COMMENT     Sequence Label: lacI-Ptac-lacO-IMBB4-pUC
FEATURES             Location/Qualifiers
     source          1..3430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     912..931
                     /label=LacI-R
                     /note="LacI, reverse primer"
     misc_feature    970..991
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     rep_origin      complement(1188..1776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1268..1287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1947..2738,2739..2807))
                     /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     2570..2589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2808..2912)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2970..2990
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     protein_bind    2999..3015
                     /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(3023..3029,3030..3046,3047..3052))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(join(3298..3430,1..950))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
       61 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
      121 tcggtatcgt cgtatcccac taccgaaata tccgcaccaa cgcgcagccc ggactcggta
      181 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
      241 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
      301 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
      361 cgcagacgcg ccgagacaga acttaatgga cccgctaaca gcgcgatttg ctggtgaccc
      421 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
      481 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
      541 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
      601 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
      661 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
      721 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
      781 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
      841 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
      901 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcat gatttatttc
      961 ctctaagtac tctagaagcg gccgcgaatt cacgcgtcaa ttgtttcttt tgggtatagc
     1021 gtcgtggaca gtcattcatc tttctgcccc tccaaaagca aaaacccgcc gaagcgggtt
     1081 tttacgtaaa tcaggtgaaa ctgaccgata agccggatcg gaaagaacat gtgagcaaaa
     1141 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1201 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1261 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1321 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1381 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1441 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1501 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1561 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1621 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1681 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1741 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1801 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1861 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1921 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1981 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2041 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2101 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2161 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2221 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2281 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2341 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2401 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2461 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2521 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2581 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2641 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2701 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2761 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2821 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2881 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     2941 gtcccaatga tatgcatgtc gacctcgagc tgcagcggcc gctactagta gtgtgaaatt
     3001 gttatccgct cacaattcca cacattatac gagccggatg attaattgtc aacagctcat
     3061 ttcagaatat ttgccagaac cgttatgatg tcggcgcaaa aaacattatc cagaacggga
     3121 gtgcgccttg agcgacacga attatgcagt gatttacgac ctgcacagcc ataccacagc
     3181 ttccgatggc tgcctgacgc cagaagcatt ggtgcaccgt gcagtcgatg ataagctgtc
     3241 aaacctctag tactaaaggc aaaatgccag cccgatcggc tggcattttt atctcaatca
     3301 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3361 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     3421 gggcaacagc
//
