LOCUS       sequence-193515-        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: MTST6
FEATURES             Location/Qualifiers
     source          1..3757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     193..210
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     194..210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     262..281
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        262..280
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1351..1367)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1351..1367)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1364..1386)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1375..1391
                     /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(1399..1405,1406..1423,1424..1429))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1444..1465
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1582..1599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1753..2341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1833..1852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2512..3303,3304..3372))
                     /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     3135..3154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3373..3477)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3545..3563
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3601..3623)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3723..3742
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
       61 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      121 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
      181 cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacctc gcgaatgcat
      241 ctagatccaa ttgaccgtac gtaatacgac tcactatagg gaatctaata acggaaccta
      301 tatggtacgt aagtatgagt agtggtttcg gtgtaaattt cagggaccaa caaggtgcta
      361 tagacgtaag gctagcttgc ctacaaatag accctaaggt atgccaccaa aggggtagta
      421 atagttactg gtggaattgg agttggctag acatgaatac gttctaccct gcttacttct
      481 acgtaataca aagttgggaa aggtggacgg actggaagta caattactac cctggtgctg
      541 taatgcctag gggtgaacct aagtggacgc cttgcaggca atgcatgcct agtcacataa
      601 atggtaggac gaataggatg caaaaggctc taggtttcca aaggagggta tactgctacc
      661 ctttcataga agctgaaatg agggaatgca tggaaagtga ccctcaccaa gacaatgtaa
      721 agtggataat attcctaaag gtaaatgtaa tatacttcat gtgggacata tacgtaaggg
      781 ctgaatgcaa tctaacgcaa cctttcatgt tctggatgaa gcaaggtttc aggttcctaa
      841 gtttcacgca cgaattcaat cacgacacgc tacactggta cctacaccaa ctaggtagtg
      901 ctcctaatgt agctggtggt tgcaggacga agagttgcca aacggactac aatttcaatc
      961 accctgacgt aaattaccct aatcaatgcg acttctgccc tttcatgcac tggctatgca
     1021 atgaagctat aaagaaggac aatagtctaa cgataaatat aaggtgcgaa caagaatgca
     1081 attggggtgg tgacgaccac gtagacgctg cttgcatacc tcaaatgtgc tacaggatgg
     1141 ctatagtaaa tacgaggagg agtcacctat acgaatacag tgtatgcaat agggaaaggt
     1201 acacgctaga aatggtacct acggctcaac ctcacataag gatagctcac ataaattggg
     1261 tattcgactt cagggtacac gtttaaacat tggatcggat cccgggcccg tcgactgcag
     1321 aggcctgcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1381 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1441 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1501 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1561 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1621 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1681 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1921 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2161 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2581 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2641 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     2701 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2941 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3001 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3181 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3361 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3421 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3481 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     3541 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     3601 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     3661 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     3721 tgcggcatca gagcagattg tactgagagt gcaccat
//
