LOCUS       sequence-248901-        3625 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 3625)
  TITLE     Dumonceaux lab phytoplasma plasmids
REFERENCE   2  (bases 1 to 3625)
  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: R018
FEATURES             Location/Qualifiers
     source          1..3625
                     /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(767..785)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(768..785)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(803..819)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(803..819)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(816..838)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    827..843
                     /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(851..857,858..875,876..881))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    896..917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1034..1051)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1205..1793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1285..1304)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1964..2755,2756..2824))
                     /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     2587..2606
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2825..2929)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3007..3462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3144..3165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3356..3375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3588..3610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3602..3619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3603..3619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggc ccgacgtcgc atgctcccgg ccgccatggc
       61 ggccgcggga attcgattga tggggcgggg gatgggacga cgacagctgt tttattaggt
      121 cagagtatga ttcgtaaagg ttttgaagct ttagacacag gtggttttaa aggggttctt
      181 ttaaaacaag gtattttaga agcatctcgt gtagtagctc aacaactttt atccaaatct
      241 cgttctgtta atactcaaga agaaatcgaa aatgttgcta ctatttcgtc tggtagctta
      301 gaaacaggaa aaattgtagc tgctgctatt gaaaaagtag gtaaaacagg agttgttaat
      361 gtggacgaat ctaagagttt tgaaactgta ttagatgttg ttgaaggtat gcaattcgaa
      421 aaaggttatg tgtctcctta ttttgtagta aatcaaaaag attctttaat agaattagaa
      481 caagctgcta ttttaattac tgatcataag attaatagta tattcgaaat taatcgtatt
      541 ttagaatatt cattaagtaa tagagttcct ttactaatta ttgctgaatc ttttgataat
      601 gaaatggtaa gtattttggt cgctaataaa aaaggtaatt acttaaatgc agttgcagtt
      661 gaagccccag gttttggcga ccaccaaatc actagtgaat tcgcggccgc ctgcaggtcg
      721 accatatggg agagctccca acgcgttgga tgcatagctt gagtattcta tagtgtcacc
      781 taaatagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
      841 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
      901 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
      961 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1021 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1081 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1141 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1201 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1261 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1321 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1381 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1441 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1501 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1561 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1621 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1681 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1741 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1801 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     1861 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     1921 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     1981 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2041 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2101 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2161 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     2221 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     2281 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     2341 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     2401 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     2461 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     2521 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     2581 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     2641 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     2701 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     2761 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     2821 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     2881 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     2941 gaaaagtgcc acctgatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     3001 atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3061 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3361 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct
     3481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     3541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     3601 ttgtaaaacg acggccagtg aattg
//
