LOCUS       sequence-310913-        3871 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Used for mycoplasma detection by qPCR. 16S insert truncated 
            internally from the strain Mycoplasma capricolum subsp. capricolum 
            strain California Kid. ATB: Carbenicilline.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3871)
  AUTHORS   Jean A, Tardy F, Allatif O, Grosjean I, Blanquier B, Gerlier D
  TITLE     Assessing mycoplasma contamination of cell cultures by qPCR using a 
            set of universal primer pairs targeting a 1.5 kb fragment of 16S 
            rRNA genes.
  JOURNAL   PLoS One. 2017 Feb 22;12(2):e0172358. doi: 
            10.1371/journal.pone.0172358. eCollection 2017.
  PUBMED    28225826
REFERENCE   2  (bases 1 to 3871)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2017-02-22"; volume: "12(2):e0172358. doi"; pages: " 
            10.1371/journal.pone.0172358. eCollection 2017"
COMMENT     Sequence Label: p_m16S
FEATURES             Location/Qualifiers
     source          1..3871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     669..685
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1633..1649)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1633..1649)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1686..1704)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1725..1741)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1725..1741)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1738..1760)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1749..1765
                     /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(1773..1779,1780..1797,1798..1803))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1818..1839
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1956..1973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2127..2715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2207..2226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2886..3677,3678..3746))
                     /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     3509..3528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3747..3851)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatgtt tgatcctggc tcaggataaa
      721 cgctggcggc atgcctaata catgcaagtc gaacgggggt gcttgcacct cagtggcgaa
      781 cgggtgagta acacgtatct aacctacctt atagcggggg ataacttttg gaaacgaaag
      841 ataataccgc atgtagatct tattatcgca tgagaaaaga tcaaaagaac cgtttggttc
      901 actatgagat ggggatgcgg cgtattagct agtaggtgag ataatagccc acctaggcga
      961 tgatacgtag ccgaactgag aggttgatcg gccacattgg gactgagata cggcccagac
     1021 tcctacggga ggcagcagta gggaattttt cacaatggac gaaagtctga tgaagcaatg
     1081 ccgcgtgagt gatgacggcc ttcgggttgt aaagctctgt tgtaagggaa gaaaaaatag
     1141 agtaggaaat gactttatct tgacagtacc ttaccagaaa gccacggcta actatgtgcc
     1201 agcagccgcg gtaatacata ggtggcaagc gttatccgga tttattgggc gtatagggtg
     1261 cgtaggcggt tttgcaagtt tgaggttaaa gtccggagct caactccggt tcgccttgaa
     1321 gactgtttta ctagaatgca agagaggtaa gcggaattcc atgtgtagcg gtgaaatgcg
     1381 tagatatatg gaagaacacc tgtggcgaaa gcggcttact ggcttgttat tgacgctgag
     1441 gcacgaaagc gtggggagca aataggatta gataccctag tagtccacgc cgtaaacgat
     1501 gagtactaag tgttggggta actcagcgct tcccaaggta ggactagcga ttggggtgaa
     1561 gtcgtaacaa ggtatccgta cgggaacgtg cggatggatc acctcctttc atcgaattcc
     1621 tgcagcccgg gggatccact agttctagag cggccgccac cgcggtggag ctccagcttt
     1681 tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct
     1741 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     1801 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1861 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1921 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1981 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2041 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2101 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2161 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2221 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2281 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2341 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2401 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2461 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2521 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     2581 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2641 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2701 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2761 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2821 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2881 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2941 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3001 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3061 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3121 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3181 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3241 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3301 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3361 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3421 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3481 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3541 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3601 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3661 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3721 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3781 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3841 ggggttccgc gcacatttcc ccgaaaagtg c
//
