LOCUS       sequence-319414-        3871 bp    DNA     circular SYN 15-DEC-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 Dec 15, 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"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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)."
     primer_bind     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     191..207
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     191..207
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1154..1170)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1155..1171)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1195..1214)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1196..1214)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1221..1238)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1221..1237)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1230..1252)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1379..1834
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1466..1485)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1676..1697
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1860..1964
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1965..2033,2034..2825)
                     /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     complement(2183..2202)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2996..3584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3485..3504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3738..3755
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag ctcgaaatta accctcacta aagggaacaa aagctggagc tccaccgcgg
      181 tggcggccgc tctagaacta gtggatcccc cgggctgcag gaattcgatg tttgatcctg
      241 gctcaggatg aacgctggcg gcatgcctaa tacatgcaag tcgaacgggg gtgcttgcac
      301 ctcagtggcg aacgggtgag taacacgtat ctaacctacc ttatagcggg ggataacttt
      361 tggaaacgaa agataatacc gcatgtagat cttattatcg catgagaaaa gatcaaaaga
      421 accgtttggt tcactatgag atggggatgc ggcgtattag ctagtaggtg agataatagc
      481 ccacctaggc gatgatacgt agccgaactg agaggttgat cggccacatt gggactgaga
      541 tacggcccag actcctacgg gaggcagcag tagggaattt ttcacaatgg acgaaagtct
      601 gatgaagcaa tgccgcgtga gtgatgacgg ccttcgggtt gtaaagctct gttgtaaggg
      661 aagaaaaaat agagtaggaa atgactttat cttgacagta ccttaccaga aagccacggc
      721 taactatgtg ccagcagccg cggtaataca taggtggcaa gcgttatccg gatttattgg
      781 gcgtataggg tgcgtaggcg gttttgcaag tttgaggtta aagtccggag ctcaactccg
      841 gttcgccttg aagactgttt tactagaatg caagagaggt aagcggaatt ccatgtgtag
      901 cggtgaaatg cgtagatata tggaagaaca cctgtggcga aagcggctta ctggcttgtt
      961 attgacgctg aggcacgaaa gcgtggggag caaataggat tagataccct agtagtccac
     1021 gccgtaaacg atgagtacta agtgttgggg taactcagcg cttcccaagg taggactagc
     1081 gattggggtg aagtcgtaac aaggtatccg tacgggaacg tgcggatgga acacctcctt
     1141 tcatcaagct tatcgatacc gtcgacctcg agggggggcc cggtacccaa ttcgccctat
     1201 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1261 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1321 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     1381 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1441 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1501 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1561 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1621 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1681 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1741 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1801 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     1861 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1921 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1981 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2041 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2101 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2161 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2221 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2281 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2341 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2401 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2461 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2521 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2581 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2641 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2701 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2761 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2821 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2881 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2941 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3001 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3061 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3121 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3181 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3241 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3301 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3361 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3421 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3481 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3541 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3601 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3661 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3721 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3781 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3841 actggaaagc gggcagtgag cgcaacgcaa t
//
