LOCUS       sequence-248899-        3617 bp    DNA     circular SYN 15-APR-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 3617)
  TITLE     Dumonceaux lab phytoplasma plasmids
REFERENCE   2  (bases 1 to 3617)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AP
FEATURES             Location/Qualifiers
     source          1..3617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      26..35
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      30..39
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(735..753)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(736..753)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(771..787)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(771..787)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(784..806)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    795..811
                     /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(819..825,826..843,844..849))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    864..885
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1002..1019)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1173..1761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1253..1272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1932..2723,2724..2792))
                     /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     2555..2574
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2793..2897)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2975..3430)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3112..3133)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3324..3343
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3556..3578
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3570..3587
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3571..3587
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3594..3613
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3594..3612
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttgggcccga cgtcgcatgc tcccggccgc catggcggcc gcgggaattc gattggcagg
       61 ggatggaacc acgactgcta ctattttagc tcaaaaaatg attcatagag gttttaaagc
      121 aatagattta ggggctaatc ctgttttagt aaaagaagga atagaaaaag caacaaaaaa
      181 agttatatca aaacttttag aaaaatcaaa accagtaata acacaagaag atatagaaaa
      241 tgtagcttct atttcatctg gaagtaaaga aattggaaaa ataattgctt tagcaatgca
      301 aaaagtaaca aaaaatggtg ttattaacgt tgatgaatct aaaggatttg aaaccaaatt
      361 agaagtagtt gaaggtatgc aatatgataa aggttacgct tcaccttatt ttgttaacaa
      421 tagggaaact atgtcagtcg atttagaaca acctttagtt ttagtaacag accataaaat
      481 aaacaatatt caagaaattt tacatttatt agaagaagta gtaaaacaag gtaagtcatt
      541 attaataatt gctgattctg ttgaaaatga agttaccagt attttagtta tcaacaaaat
      601 gaaaggaaca tttaatatag ttgttactaa tgcccctggt tttggcgaca atcaaatcac
      661 tagtgaattc gcggccgcct gcaggtcgac catatgggag agctcccaac gcgttggatg
      721 catagcttga gtattctata gtgtcaccta aatagcttgg cgtaatcatg gtcatagctg
      781 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      841 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      901 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      961 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1021 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1081 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1141 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1201 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1261 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1321 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1381 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1441 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1501 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1561 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1621 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1681 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1741 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1801 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1861 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1921 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1981 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2041 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2101 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2161 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2221 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2281 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2341 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2401 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2461 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2521 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2581 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2641 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2701 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2761 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2821 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2881 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgatgcggt gtgaaatacc
     2941 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     3001 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3061 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3121 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3181 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3241 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3301 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3361 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3421 cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3481 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     3541 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg
     3601 actcactata gggcgaa
//
