LOCUS       sequence-248861-        3609 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 3609)
  TITLE     Dumonceaux lab phytoplasma plasmids
REFERENCE   2  (bases 1 to 3609)
  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: PYLR
FEATURES             Location/Qualifiers
     source          1..3609
                     /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(774..792)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(775..792)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(810..826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(810..826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(823..845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    834..850
                     /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(858..864,865..882,883..888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    903..924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1041..1058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1212..1800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1292..1311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1971..2762,2763..2831))
                     /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     2594..2613
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2832..2936)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3014..3469)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3151..3172)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3363..3382
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(3595..3609,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3609,1..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 tgatggggca ggggatggaa
      121 caacgactgc tactctttta gctcaaaaaa tgattcatcg tggtttcaaa gcaatagatt
      181 taggtgctaa tcctgtttta gtaaaagaag gaatagaaaa agcgacaaaa caagttatag
      241 caaaactttt agaaaaatca aaaccagtaa taacacaaga agatatcgaa aatgtagctt
      301 ctatttcttc tggaagtaaa gaaattggaa aaataattgc tttagcaatg caaaaagtaa
      361 ccaaaaatgg tgttattaat gttgacgaat caaaaggatt tgaaactaaa ttagaagtag
      421 ttgaaggtat gcaatatgac aaaggttatg cttcgcctta ttttgttaat aatagagaaa
      481 ctatgtcagt tgaattagaa caacctttag ttttagtaac agatcataaa ataaataata
      541 ttcaagaaat tttacattta ttagaagaag tagtgaaaca aggtaagtca ttattaataa
      601 ttgctgattc tgtagaaaat gaagttacta gtattttagt tatcaacaaa atgaaaggga
      661 catttaatat agttgttact aatgcccctg gttaatcact agtgaattcg cggccgcctg
      721 caggtcgacc atatgggaga gctcccaacg cgttggatgc atagcttgag tattctatag
      781 tgtcacctaa atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
      841 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
      901 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
      961 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1021 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1081 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1141 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1201 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1261 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1321 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1381 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1441 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1501 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1561 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1621 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1681 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1741 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1801 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1861 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1921 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     1981 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2041 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2101 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2161 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2221 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     2281 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     2341 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2401 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2461 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2521 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2581 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2641 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2701 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2761 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     2821 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2881 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     2941 tttccccgaa aagtgccacc tgatgcggtg tgaaataccg cacagatgcg taaggagaaa
     3001 ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3061 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3121 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3181 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3241 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3301 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3361 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3421 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat
     3481 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     3541 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     3601 cacgacgtt
//
