LOCUS       sequence-248864-        3622 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 3622)
  TITLE     Dumonceaux lab phytoplasma plasmids
REFERENCE   2  (bases 1 to 3622)
  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: PGY
FEATURES             Location/Qualifiers
     source          1..3622
                     /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(787..805)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(788..805)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(823..839)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(823..839)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(836..858)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    847..863
                     /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(871..877,878..895,896..901))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    916..937
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1054..1071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1225..1813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1305..1324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1984..2775,2776..2844))
                     /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     2607..2626
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2845..2949)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3027..3482)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3164..3185)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3376..3395
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(3608..3622,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3622,1..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 tgatggggcg ggggatggga
      121 cgacgacagc aattgtttta gctcaaagta tgattcaaaa aggttttaaa tttgttaatt
      181 ctggtgctca atctgttcta gttaaaaaag gaattcttaa agcttcacaa aaagtcatag
      241 aacaaatttt agaaaaatct aaaccaattt ctacacaaga agagatagct aatatcgcta
      301 cactttcttc aggtagtaaa gaaataggcg aaattatagt ttctgcaata aataaagtta
      361 ctcaaaaagg aataattagc ataggagaat ctaaaggttt agaaactgaa ttagaagttg
      421 ttgaaggaat gcaatatgat aaaggttatt tatcaagtat tttcgttaat aaattggcca
      481 atatgtcagt cgaattcgaa agagcattaa ttttggtaac tgatcataaa attaataaca
      541 ttaacgaaat taatcactta ttggaagaag ttaaagcgaa aactcaacct ttgttaatta
      601 tagctaattc gtttgataac gatgtaatta atattttggc gctaaataag tttcatggta
      661 ttttaaatat cgctgctact gatgccccta gttttggcga ccaccaaatc actagtgaat
      721 tcgcggccgc ctgcaggtcg accatatggg agagctccca acgcgttgga tgcatagctt
      781 gagtattcta tagtgtcacc taaatagctt ggcgtaatca tggtcatagc tgtttcctgt
      841 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
      901 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
      961 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1021 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1081 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1141 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1201 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1261 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1321 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1381 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1441 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1501 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1561 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1621 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     1681 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1741 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     1801 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     1861 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     1921 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     1981 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2041 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2101 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2161 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2221 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2281 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     2341 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     2401 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     2461 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     2521 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     2581 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     2641 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     2701 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2761 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     2821 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     2881 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2941 ggttccgcgc acatttcccc gaaaagtgcc acctgatgcg gtgtgaaata ccgcacagat
     3001 gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt taaaattcgc
     3061 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3121 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3181 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3241 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3301 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3361 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3421 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3481 gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     3541 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     3601 gggttttccc agtcacgacg tt
//
