LOCUS       sequence-254611-        2481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad-host-range toolkit (BTK) entry vector, with superfolder GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2481)
  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 2481)
  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: pBTK001
FEATURES             Location/Qualifiers
     source          1..2481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..12
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             19..732
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      744..815
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      831..858
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(882..916)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(987..1646)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1561..1580
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(1752..2297)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1832..1851)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaagaggaga aatactagat gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt
       61 ctggtggaac tggatggtga tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa
      121 ggtgacgcaa ctaatggtaa actgacgctg aagttcatct gtactactgg taaactgccg
      181 gttccttggc cgactctggt aacgacgctg acttatggtg ttcagtgctt tgctcgttat
      241 ccggaccata tgaagcagca tgacttcttc aagtccgcca tgccggaagg ctatgtgcag
      301 gaacgcacga tttcctttaa ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt
      361 gaaggcgata ccctggtaaa ccgcattgag ctgaaaggca ttgactttaa agaggacggc
      421 aatatcctgg gccataagct ggaatacaat tttaacagcc acaatgttta catcaccgcc
      481 gataaacaaa aaaatggcat taaagcgaat tttaaaattc gccacaacgt ggaggatggc
      541 agcgtgcagc tggctgatca ctaccagcaa aacactccaa tcggtgatgg tcctgttctg
      601 ctgccagaca atcactatct gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa
      661 cgcgatcata tggttctgct ggagttcgta accgcagcgg gcatcacgca tggtatggat
      721 gaactgtaca aatgaccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
      781 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
      841 cgggtgggcc tttctgcgtt tatacgtctc tgaccagacc aataaaaaac gcccggcggc
      901 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
      961 ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt
     1021 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     1081 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     1141 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     1201 ggattggctg aaacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     1261 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     1321 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     1381 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga
     1441 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     1501 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     1561 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     1621 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     1681 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     1741 tgcccgatca atttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
     1801 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     1861 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     1921 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     1981 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     2041 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     2101 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     2161 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     2221 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     2281 gaccaaaacg atctcaaccc ctgattctgt ggataaccgt agtcggcgag acggaaagtg
     2341 aaacgtgatt tcatgcgtca ttttgaacat tttgtaaatc ttatttaata atgtgtgcgg
     2401 caattcacat ttaatttatg aatgttttct taacatcgcg gcaactcaag aaacggcagg
     2461 ttcggatctt agctactaga g
//
