LOCUS       sequence-431081-        3214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pUCIDT-AMP containing 16S rRNA gene of F. prausnitzii for absolute 
            concentration quantification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3214)
  AUTHORS   Maghini DG, Dvorak M, Dahlen A, Roos M, Kuersten S, Bhatt AS
  TITLE     Quantifying bias introduced by sample collection in relative and 
            absolute microbiome measurements.
  JOURNAL   Nat Biotechnol. 2024 Feb;42(2):328-338. doi: 
            10.1038/s41587-023-01754-3. Epub 2023 Apr 27.
  PUBMED    37106038
REFERENCE   2  (bases 1 to 3214)
  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"; doi: 
            "10.1038/s41587-023-01754-3"; journalName: "Nat Biotechnol"; date: 
            "2024-02"; volume: "42"; issue: "2"; pages: "328-338"
COMMENT     Sequence Label: pUCIDT-AMP-Fp16SrRNA
FEATURES             Location/Qualifiers
     source          1..3214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(993..1009)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(993..1009)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1006..1028)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1017..1033
                     /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(1041..1047,1048..1065,1066..1071))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1086..1107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1224..1241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1395..1983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1475..1494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2154..2945,2946..3014))
                     /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     2777..2796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3015..3119)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3187..3205
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accaaatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcatcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgcaac gcgatgacga tggatagcga
      421 ttcatcgatg agctgacccg atcgccgccg ccggagggtt gcgtttgaga cgggcgacag
      481 atgcccagac tcctacggga ggcagcagtg gggaatattg cacaatgggg gaaaccctga
      541 tgcagcgacg ccgcgtggag gaagaaggtc ttcggattgt aaactcctgt tgttgaggaa
      601 gataatgacg gtactcaaca aggaagtgac ggctaactac gtgccagcag ccgcggtaaa
      661 acgtaggtca caagcgttgt ccggaattac tgggtgtaaa gggagcgcag gcgggaaggc
      721 aagttggaag tgaaatccat gggctcaacc catgaactgc tttcaaaact gtttttcttg
      781 agtagtgcag aggtaggcgg aattcccggt gtagcggtgg aatgcgtaga tatcgggagg
      841 aacaccagtg gcgaaggcgg cctactgggc accaactgac gctgaggctc gaaagtgtgg
      901 gtagcaaaca ggattagata ccctggtagt ccacactgtg gccgatcagt tctggaccag
      961 cgagctgtgc tgcgactcgt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1021 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1081 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1141 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1201 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1261 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1321 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1381 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1441 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1501 agctccctcg tgcgctctcc tgttccgacc ctgtcgctta ccggatacct gtccgccttt
     1561 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1621 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1681 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1741 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1801 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1861 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1921 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1981 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2041 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2101 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2161 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2221 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     2281 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     2341 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     2401 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2461 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2521 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2581 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2641 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2701 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2761 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2821 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2881 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2941 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3001 tgttgaatac tcatactcta cctttttcaa tattattgaa gcatttatca gggttattgt
     3061 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3121 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     3181 tataaaaata ggcgtatcac gaggcccttt cgtc
//
