LOCUS       sequence-193516-        3757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Custom designed synthetic DNA fragment lacking homology to natural 
            sequences for preparation of internal standards for RNA-seq in 
            metatranscriptomics..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3757)
  AUTHORS   Satinsky BM, Gifford SM, Crump BC, Moran MA
  TITLE     Use of internal standards for quantitative metatranscriptome and 
            metagenome analysis.
  JOURNAL   Methods Enzymol. 2013;531:237-50. doi: 
            10.1016/B978-0-12-407863-5.00012-5.
  PUBMED    24060124
REFERENCE   2  (bases 1 to 3757)
  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: "Methods 
            Enzymol."; date: "2013"; volume: "531:237-50. doi"; pages: " 
            10.1016/B978-0-12-407863-5.00012-5"
COMMENT     Sequence Label: MTST5
FEATURES             Location/Qualifiers
     source          1..3757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..30
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     224..246
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     238..255
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     239..255
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     307..326
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        307..325
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1396..1412)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1396..1412)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1409..1431)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1420..1436
                     /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(1444..1450,1451..1468,1469..1474))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1489..1510
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1627..1644)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1798..2386)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1878..1897)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2557..3348,3349..3417))
                     /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     3180..3199
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3418..3522)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3590..3608
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3646..3668)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
       61 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
      121 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
      181 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
      241 aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa tgcatctaga tccaattgac
      301 cgtacgtaat acgactcact atagggttcg gtggtctata ctactaccta agttggatgt
      361 actggtggaa gtgctaccaa cacagtaatg ctggtatagg taggcaacac tacgacttca
      421 ggaagagtct aacgaatgta tgcataatac taatgcctta catgtggaag caccctataa
      481 cggacaggat gaggcacagg cacatgtgca ggcaagcttt caagtggatg tgcggtgaaa
      541 ataagttctg gctagtaagg gctatggaaa atcaacctga cgaaaggata ctagctcaaa
      601 tgacgataac ggacagtgac tggcaacctg aagaatggta caagaagagg cacgaccctg
      661 gtgaaaatga cgtaataagg tgcgtataca taggtataga aaatctagta aatgctacgt
      721 gccctgacat ggaagactac tacgctatga cgggtaataa gcctctacta gaactaaata
      781 gtataggtcc ttgcacgcaa tgcacggtac acaagctaga aggtgtacac tgcatatggt
      841 ggatagtaag gagggaccac ttccctgtac ctataataca aatagtagac gtattcaatc
      901 tatacaattt cgctagtggt acggtactat gcatacaaca cgctgctcac ccttggggtg
      961 actggatgtt cgacgtacaa tacgaaagtt gcaggatgta caggtggtgg atgacgagga
     1021 atgactggag tggtcctaat aagtggagtg gtgctcacag tatatgccaa cctcactgct
     1081 gcaggagtga cgacagtagg gtaagtaaga ggatggctac ggtaacgaag gaagtagtag
     1141 aaatgagtca catggaccta aagaggagtg cttactgcaa taggacgcaa ctagaagaat
     1201 acgacgcttt ctacacgagg tggaagttcg taccttggat gtaccctgct cctctacctt
     1261 gcgaagtaca agacttcgta acgaggagga cgttcgacta ccctgaccct acggcttgcg
     1321 gtctagttta aacattggat cggatcccgg gcccgtcgac tgcagaggcc tgcatgcaag
     1381 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1441 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1501 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1561 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1621 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1681 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1741 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1801 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1861 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1921 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1981 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2041 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2101 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2161 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2221 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2281 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2341 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2401 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2461 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2521 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2581 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2641 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2701 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2761 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2821 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2881 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2941 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3001 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3061 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3121 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3181 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3241 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3301 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3361 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3421 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3481 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3541 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3601 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3661 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3721 gggcgcgtca gcgggtgttg gcgggtgtcg gggctgg
//
