LOCUS       sequence-431080-        3232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pUCIDT-AMP containing 16S rRNA gene of B. vulgatus for absolute 
            concentration quantification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3232)
  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 3232)
  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-Bv16SrRNA
FEATURES             Location/Qualifiers
     source          1..3232
                     /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(1011..1027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1011..1027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1024..1046)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1035..1051
                     /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(1059..1065,1066..1083,1084..1089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1104..1125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1242..1259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1413..2001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1493..1512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2172..2963,2964..3032))
                     /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     2795..2814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3033..3137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3205..3223
                     /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 atggtccaaa ctcctacggg aggcagcagt gaggaatatt ggtcaatggg cgagagcctg
      541 aaccagccaa gtagcgtgaa ggatgactgc cctatgggtt gtaaacttct tttataaagg
      601 aataaagtcg ggtatggata cccgtttgca tgtactttat gaataaggat cggctaactc
      661 cgtgccagca gccgcggtaa tacggaggat ccgagcgtta tccggattta ttgggtttaa
      721 agggagcgta gatggatgtt taagtcagtt gtgaaagttt gcggctcaac cgtaaaattg
      781 cagttgatac tggatatctt gagtgcagtt gaggcaggcg gaattcgtgg tgtagcggtg
      841 aaatgcttag atatcacgaa gaactccgat tgcgaaggca gcctgctaag ctgcaactga
      901 cattgaggct cgaaagtgtg ggtatcaaac aggattagat accctggtag tccacacggt
      961 aaatcagttc tggaccagcg agctgtgctg cgactcgtgg cgtaatcatg gtcatagctg
     1021 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1081 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1141 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1201 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1261 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1321 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1381 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1441 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1501 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gtcgcttacc
     1561 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1621 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1681 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1741 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1801 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1861 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1921 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1981 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2041 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2101 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2161 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2221 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2281 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2341 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2401 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2461 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2521 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2581 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2641 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2701 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2761 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2821 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2881 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2941 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3001 ataagggcga cacggaaatg ttgaatactc atactctacc tttttcaata ttattgaagc
     3061 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3121 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     3181 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
