LOCUS       sequence-248898-        3621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains the chaperonin-60 universal target from phytoplasma 
            baceria; used as a control for development of molecular diagnostic 
            assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3621)
  TITLE     Dumonceaux lab phytoplasma plasmids
REFERENCE   2  (bases 1 to 3621)
  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"
COMMENT     Sequence Label: FD
FEATURES             Location/Qualifiers
     source          1..3621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      73..82
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      77..86
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(786..804)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(787..804)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(822..838)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(822..838)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(835..857)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    846..862
                     /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(870..876,877..894,895..900))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    915..936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1053..1070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1224..1812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1304..1323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1983..2774,2775..2843))
                     /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     2606..2625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2844..2948)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3026..3481)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3163..3184)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3375..3394
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(3607..3621,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3621,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg ggcccgacgt
       61 cgcatgctcc cggccgccat ggcggccgcg ggaattcgat tatggggcgg gggatgggac
      121 gacgacagca attgttttag ctcaaagtat gattcaaaaa ggttttaaat ttgttaattc
      181 tggtgctcaa tctgttctag ttaaaaaagg aattcttaaa gcttcacaaa aagtcataga
      241 acaaatttta gaaaaatcta aaccaatttc tacacaagaa gaaatagcta atatcgctac
      301 actttcttca ggtagtaaag aaataggtga aattatagtt tctgcaataa ataaagttac
      361 tcaaaaagga ataattagca taggagaatc taaaggttta gaaactgaat tagaagttgt
      421 tgaaggaatg caatatgata aaggttattt atcaagtatt ttcgttaata aattagccaa
      481 tatgtcagtc gaattcgaaa gagcattaat tttggtaact gaccataaaa ttaataacat
      541 taacgaaatt aataatttat tggaagaagt taaagcgaaa actcaacctt tgttaattat
      601 agctaattcg tttgataacg atgtaattaa tattttggct ctaaataagt ttcatggtat
      661 tttaaatatc gctgctactg aagcccctgg ttttggcgac aaccaaatca ctagtgaatt
      721 cgcggccgcc tgcaggtcga ccatatggga gagctcccaa cgcgttggat gcatagcttg
      781 agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg
      841 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      901 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      961 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1021 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1621 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1681 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1741 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1801 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1861 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1921 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1981 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2041 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2101 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2161 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2221 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     2281 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2341 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2401 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2461 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2521 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2581 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2641 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2701 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2761 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2821 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2881 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2941 gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg
     3001 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg
     3061 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3121 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3181 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3241 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3301 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3361 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3421 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3481 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     3541 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     3601 ggttttccca gtcacgacgt t
//
