LOCUS       sequence-248869-        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: Cr
FEATURES             Location/Qualifiers
     source          1..3621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      50..59
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      54..63
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(763..781)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(764..781)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(799..815)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(799..815)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(812..834)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    823..839
                     /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(847..853,854..871,872..877))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    892..913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1030..1047)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1201..1789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1281..1300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1960..2751,2752..2820))
                     /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     2583..2602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2821..2925)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3003..3458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3140..3161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3352..3371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3584..3606
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3598..3615
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3599..3615
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggc ccgacgtcgc atgctcccgg ccgccatggc
       61 ggccgcggga attcgattga tgggtcgggg gacgggacga cgacagctac agtattagca
      121 caaaaaatga ttcataaagg atttcagttt gttaattctg gtgctaaagc tgtttctatt
      181 aaagaaggta ttatgagagc ttcccaagaa gtagtatcaa aactgttagc taaatcaaaa
      241 cctgtcgaaa cacaaagtga tattgaaaac gtagcaacta tttcttctgg tcaaaaagat
      301 ataggccgta ttatagcttc ggcaatggaa aaagttacta aaaaaggtgt tattaacgta
      361 gatgaatcta aaggatttga aactgaactt gaagtagtaa aaggtttgaa atatgataaa
      421 ggttatattt ctcctcattt tgtcaccaac aaagaaaata tgtctgtaga atttgaacaa
      481 gcttcagtat tagttacaga tcataaaatt agtaatttac aagaaattcg ttctatttta
      541 gaagatgtag taaaaaattc tactccttta cttattattg ctgcttcttt tgaaaatgac
      601 gttataggtg ctttagtttt taataaaatt agcggagtat ttaatgtagt tgctactgat
      661 gcccctggtt ttggcgacaa caaatcacta gtgaattcgc ggccgcctgc aggtcgacca
      721 tatgggagag ctcccaacgc gttggatgca tagcttgagt attctatagt gtcacctaaa
      781 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
      841 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
      901 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
      961 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1021 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1081 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1141 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1201 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1261 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1321 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1381 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1441 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1501 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1561 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1621 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1681 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1741 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1801 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1861 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     1921 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     1981 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2041 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2101 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2161 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2221 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2281 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2341 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2401 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     2461 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     2521 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     2581 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     2641 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     2701 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     2761 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     2821 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     2881 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     2941 agtgccacct gatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     3001 ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3061 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc
     3481 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     3541 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     3601 aaaacgacgg ccagtgaatt g
//
