LOCUS       sequence-248870-        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: RS
FEATURES             Location/Qualifiers
     source          1..3621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..26
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      56..65
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      60..69
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(769..787)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(770..787)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(805..821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(805..821)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(818..840)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    829..845
                     /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(853..859,860..877,878..883))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    898..919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1036..1053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1207..1795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1287..1306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1966..2757,2758..2826))
                     /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     2589..2608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2827..2931)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3009..3464)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3146..3167)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3358..3377
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3590..3612
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3604..3621
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3605..3621
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattgtaat acgactcact atagggcgaa ttgggcccga cgtcgcatgc tcccggccgc
       61 catggcggcc gcgggaattc gattgatggg tcgggggatg ggacgacgac agcaattgtt
      121 ttagctcaaa gtatgattca aaaaggtttt aaatttgtta attctggtgc tcaatctgtt
      181 ctagttaaaa aaggaattct taaagcttca caaaaagtca tagaacaaat tttagaaaaa
      241 tctaaaccaa tttctacaca agaagaaata gctaatatcg ctatactttc ttcaggtagt
      301 aaagaaatag gagaaattat agtttctgca ataaataaag ttactcaaaa aggaataatt
      361 agcataggag aatctaaagg tttagaaact gaattagaag ttgttgaagg aatgcaatat
      421 gataaaggtt atttatcaag tattttcgtt aataaattgg ccaatatgtc agtcgaattc
      481 gaaagagcat taattttggt aactgatcat aaaattaata acattaatga aattaatcat
      541 ttattggaag aagttaaagc aaaaactcaa tctttgttaa ttatagctaa ttcgtttgat
      601 aacgatgtaa ttaatatttt ggcgctaaat aagtttcatg gtattttaaa tattgctgct
      661 actgaagccc caagttttgg cgacaaccaa tcactagtga attcgcggcc gcctgcaggt
      721 cgaccatatg ggagagctcc caacgcgttg gatgcatagc ttgagtattc tatagtgtca
      781 cctaaatagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
      841 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
      901 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
      961 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1021 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1081 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1141 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1621 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1741 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1801 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1861 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1921 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1981 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2041 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2101 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2161 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2221 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2281 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2341 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2401 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2461 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2521 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2581 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2641 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2701 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2761 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2821 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2881 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2941 ccgaaaagtg ccacctgatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3001 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3061 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3121 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3181 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3241 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3301 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3361 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3421 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg
     3481 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     3541 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     3601 cgttgtaaaa cgacggccag t
//
