LOCUS       sequence-324340-        1742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BTK Type 2 promoter for golden gate assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1742)
  AUTHORS   Leonard SP, Perutka J, Powell JE, Geng P, Richhart DD, Byrom M, Kar 
            S, Davies BW, Ellington AD, Moran NA, Barrick JE
  TITLE     Genetic Engineering of Bee Gut Microbiome Bacteria with a Toolkit 
            for Modular Assembly of Broad-Host-Range Plasmids.
  JOURNAL   ACS Synth Biol. 2018 May 18;7(5):1279-1290. doi: 
            10.1021/acssynbio.7b00399. Epub 2018 Apr 13.
  PUBMED    29608282
REFERENCE   2  (bases 1 to 1742)
  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.7b00399"; journalName: "ACS Synth Biol"; date: 
            "2018-05-18- 18"; volume: "7"; issue: "5"; pages: "1279-1290"
COMMENT     Sequence Label: pBTK103
FEATURES             Location/Qualifiers
     source          1..1742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     139..158
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(353..358,359..376,377..383)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    391..407
                     /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)."
     RBS             412..423
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(446..480)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(551..1210)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1125..1144
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(1392..1742,1..238)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
       61 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      121 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      181 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      241 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      301 ctttcctgcg ttatcccctg attctgtgga taaccgtagt cggtctcaaa cgtttacact
      361 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt taaagaggag
      421 aaattaacta tgtgagacca gaccaataaa aaacgcccgg cggcaaccga gcgttctgaa
      481 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
      541 cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca
      601 ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca
      661 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt
      721 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgaaacga
      781 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
      841 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
      901 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
      961 tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg
     1021 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa
     1081 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     1141 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     1201 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     1261 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgcccg atcaatcatg
     1321 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1381 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1441 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1501 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     1561 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1621 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1681 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1741 ga
//
