LOCUS       sequence-248857-        3615 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 3615)
  TITLE     Dumonceaux lab phytoplasma plasmids
REFERENCE   2  (bases 1 to 3615)
  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: SbGP-1
FEATURES             Location/Qualifiers
     source          1..3615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..10
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5..14
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(708..726)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(709..726)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(744..760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(744..760)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(757..779)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    768..784
                     /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(792..798,799..816,817..822))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    837..858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(975..992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1146..1734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1226..1245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1905..2696,2697..2765))
                     /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     2528..2547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2766..2870)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2948..3403)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3085..3106)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3297..3316
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3529..3551
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3543..3560
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3544..3560
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3567..3586
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3567..3585
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gccgccatgg cggccgcggg aattcgattc gggggatggg acgacgacag caactgtttt
       61 agctcaaagt atgattcatc gtggttttaa agcaattgat agaggggcta atcctatttt
      121 cgttaaagaa gggatcgaat tagcgtctca agtggtttct caaaagctat tggctaaaac
      181 taaaaaagta gcagctcaag aggatatcca aaatgtagca gctatttctt caggaagtca
      241 agaaattggt aaaattattg ctcaagctat gaaaaaagtt ggtaaaaatg gtgttattaa
      301 tgttgatgaa tcaaaaggat ttgaaaccga gttagaagta gttcaaggtt tacaatatga
      361 taaaggatat gcttctccgt attttgcttc agatagagaa tctatggcgg ttgaattaaa
      421 ccaaccatta gttttagtga ccgatcataa aattagtacc attcaagaaa ttttgcctat
      481 tttagaaggt gttgttaaag aatctaaatc tttattaatt gtagctgaat cagttgaaaa
      541 tgaagtttta ggggttttgg ttgctaataa attaaggggt acttttaatg tagttgttac
      601 tgaagcccct ggttttggcg accaccaaat cactagtgaa ttcgcggccg cctgcaggtc
      661 gaccatatgg gagagctccc aacgcgttgg atgcatagct tgagtattct atagtgtcac
      721 ctaaatagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
      781 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
      841 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
      901 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
      961 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1021 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1081 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1141 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1201 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1261 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1321 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1381 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1441 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1501 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1561 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1621 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1681 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1741 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1801 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1861 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     1921 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1981 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2041 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2101 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2161 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2221 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2281 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2341 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2401 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2461 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2521 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2581 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2641 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     2701 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     2761 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     2821 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     2881 cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     2941 catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3001 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3061 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3121 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3181 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3241 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3301 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3361 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc
     3421 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     3481 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     3541 gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa ttgggcccga
     3601 cgtcgcatgc tcccg
//
