LOCUS       sequence-491211-        5870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (Methanomassiliicoccus luminyensis B10) 16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5870)
  AUTHORS   Van Beek JM, Robles G, Mewalal R, Blaby I, Hatzenpichler RO
  TITLE     A collection of archaeal 16S rRNA Clone-FISH cultures for probe 
            validation in fluorescence in situ hybridization experiments
  JOURNAL   Microbiol Resour Announc. 2025 October. 0:e01131-25.
REFERENCE   2  (bases 1 to 5870)
  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: "Microbiol 
            Resour Announc. 2025 October. 0:e01131-25."
COMMENT     Sequence Label: MlumB10
FEATURES             Location/Qualifiers
     source          1..5870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(380..427)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1903..1922)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1904..1922)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1929..1946)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1929..1945)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1938..1960)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2187..2296
                     /note="incP origin of transfer"
     rep_origin      2486..2914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2573..2592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2783..2804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3258..4052
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3312..3331)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3922..3941
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4070..4138,4139..4930)
                     /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(4288..4307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5101..5689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5590..5609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5843..5860
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgg cttaagccga
      301 attctgcaga tatccatcac actggcggcc gctcgagcat gcatctagac catcacactg
      361 gcggccgctc gagcatgcac aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
      421 atgctagtag gaggtgatcc atctgcaggt tcccctacag ataccttgtt acgacttaac
      481 cctccttgct gaatctcagt tcgaaagccc caataagaaa catcctcact gaaacccaac
      541 tcgggtggtt tgacgggcgg tgtgtgcaag gagcaggggc atattcaccg cttgatgttg
      601 acaagcgatt actacggaat ccagcttcat gggggcgggt tacagccccc aatccgaact
      661 atggacgggt ttcaggattg tctccacctt tcggtgtcga agcccattgt cccgtccttt
      721 gtagcgcgcg tgttgcccag gggattcggg gcatacggac ctaccgttga cctctccttc
      781 ctctgactta gcgccagcgg tcccattaat gtgcgccccc tccggagaag ggactagcaa
      841 ttaatggtga gggtctcgtt cgttatctca cttaagagaa cgccttacgg tacgaactga
      901 cgacggccat gcaccacctc tcggaaaatc aggtaaagtc atcggcttga ccttcatata
      961 tccgtcgctc ctggtgagtt tttcggcgtt gaatccaatt gaaccgcacg ctcctcccgt
     1021 tgcggtgctc ccccgccaat tcctttaagt ttcatccttg cggacatact ccccaagcag
     1081 caggcttaac aacttctctc cggcactggg cgcccactaa ggacccccaa caccaagccc
     1141 gcagcgttta caccctggac taccggggta tctaatcccg tttgcgcccc agggcttcgt
     1201 ccctcaccgt cggatccgtt ctagtcagac gccttcgcca ccggtggtcc ttccaggatt
     1261 acaggatttt acccctaccc cagaagtacc tctgacctct cccggtccca agtctcccag
     1321 tctccccgga attcggacag ttgagctgcc cgatttaccc aaggatttag aagaccggct
     1381 acggacgttt taggctcaat aatatcgacc accacttggg gcgcgggtat taccgcggcg
     1441 gctggcaccc gtcttacccg ccccttactt ctcctgtttt ttagacagaa gaacagccaa
     1501 cacaaagtgc tggcacttgg aattccctcg tcggggtttc ccccattgcg aagttttcgc
     1561 gactgctgcg ccccgtaggg cctggactca tgtctcagag tccatctctg ggctccctct
     1621 ctcaaggccc atatccgtca cgggctagtg ggtccattac acccactact gcctgataga
     1681 ccgcagacca atcctaaggc accggagttt tcggcctagt agcattccag catgctagac
     1741 ctatggggta ttatcctcag tttcccgaga ttatccccca ccttagggca tgttatccac
     1801 gtgttactga gcggtctgcc gagaccttac gtctctcgac tcgcatgtct taatcgtatt
     1861 ccaatagcgg tggccgccgg caggatcaac cggaataatt cgccctatag tgagtcgtat
     1921 tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     1981 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2041 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ctggaaggca
     2101 gtacaccttg ataggtgggc tgcccttcct ggttggcttg gtttcatcag ccatccgctt
     2161 gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag
     2221 caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac
     2281 ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct
     2341 ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat
     2401 gaccccgaag cagggttatg cagcggaaaa gcgctgcttc cctgctgttt tgtggaatat
     2461 ctaccgactg gaaacaggca aatgcaggaa ccctgtagcg gcgcattaag cgcggcgggt
     2521 gtggtggtta cgcgcagcgt gaccgctaca cttgctagcg ccctagcgcc cgctcctttc
     2581 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2641 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2701 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2761 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2821 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2881 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaattcaggg cgcaagggct
     2941 gctaaaggaa gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga
     3001 atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag
     3061 cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac
     3121 cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga
     3181 tggctttctt gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag
     3241 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3301 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3361 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3421 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3481 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3541 gcgaagtgcc ggggcaggat ctcctgtcat cccaccttgc tcctgccgag aaagtatcca
     3601 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3661 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3721 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3781 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3841 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3901 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3961 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4021 ccttctatcg ccttcttgac gagttcttct gaattgaaaa aggaagagta tgagtattca
     4081 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4141 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4201 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4261 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4321 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4381 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4441 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4501 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4561 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4621 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4681 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4741 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4801 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4861 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4921 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4981 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5041 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5101 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5161 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5221 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5281 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5341 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5401 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5461 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5521 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5581 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5641 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5701 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5761 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5821 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
