LOCUS       sequence-491213-        5440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (13cm_4210_3138-4177(-) _Thor) 16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5440)
  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 5440)
  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: Thor-LSSM13-A
FEATURES             Location/Qualifiers
     source          1..5440
                     /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(1473..1492)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1474..1492)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1499..1516)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1499..1515)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1508..1530)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            1757..1866
                     /note="incP origin of transfer"
     rep_origin      2056..2484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2143..2162)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2353..2374
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2828..3622
                     /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(2882..2901)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3492..3511
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3640..3708,3709..4500)
                     /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(3858..3877)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4671..5259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5160..5179
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5413..5430
                     /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 atgctaggga cgacacctca ccccaaggcc gctcgcatgg agcgacgggc ggtgtgtgca
      481 agaagcaggg acgtattctc cgcgccatag tgagacgcgg atactaggga tctcctgtcc
      541 acattcgaag tgaatgtgaa atccaggttc acgcgggtga gttccaaccc gcgatcccga
      601 ctacgactgg gtttcgcgat tgacttcacc tttcggtgtt gtgacgcatt gtcccagcca
      661 ttgtagcacg cgtgtggccc cgcccatatc gcggcatact gacctaccgt ggcccccacc
      721 ttcctccagc ttttcgctgg cggtccccct agtgtgcccc gtcagggccg aagcccccgg
      781 ctggcaacta ggtggcaggg atctcattcg ttgcagcact taagctgata cctcacggca
      841 cgaatttacg atggccatgc accagctctc agctcgtcga gcaagatcat cagtctggct
      901 ctcattcggc tgtctgggcg ggtaagattt ccggcgttga gtccaattaa accgcaagct
      961 ccaccccttg tggtgctccc ccgccaattc ctttaagtct cctgaggaga gtcggaatgg
     1021 ctctctaatt cagtcttgcg accgtacttc ccaagcggtc cgtttatcgc tttcgctaca
     1081 gcaccgcagt ggctcgtggc caatgcaaca cttaacggac atcgtttacg cccaggacta
     1141 ctcgggtatc taatccgatt tgctcccctg ggtttcgtcc ctcaccgtcg ggcacgttct
     1201 cgacgaccgc cttcgccact ggtggtcctc cgtagatcat tagatttcac ccttacctac
     1261 ggaataccgt cgccgtctcc cgtccccaag taaagcggta tccctaccag actgaccgtt
     1321 gagcggccag atttaagcag ggacctacag aaccggctac gaacgcttta agctcaataa
     1381 tcatggacac cactcgaaga gctggtttta ccgcggcggc tggcaccagc cttgccctct
     1441 cctttctagc ggtccacctt acggataatt cgccctatag tgagtcgtat tacaattcac
     1501 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     1561 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     1621 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ctggaaggca gtacaccttg
     1681 ataggtgggc tgcccttcct ggttggcttg gtttcatcag ccatccgctt gccctcatct
     1741 gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag caccgccagg
     1801 tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc
     1861 ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat
     1921 cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag
     1981 cagggttatg cagcggaaaa gcgctgcttc cctgctgttt tgtggaatat ctaccgactg
     2041 gaaacaggca aatgcaggaa ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2101 cgcgcagcgt gaccgctaca cttgctagcg ccctagcgcc cgctcctttc gctttcttcc
     2161 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2221 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2281 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2341 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2401 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2461 tttaacaaaa atttaacgcg aattttaaca aaattcaggg cgcaagggct gctaaaggaa
     2521 gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta
     2581 ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg
     2641 gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc
     2701 agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt
     2761 gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc
     2821 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     2881 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     2941 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3001 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3061 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3121 ggggcaggat ctcctgtcat cccaccttgc tcctgccgag aaagtatcca tcatggctga
     3181 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3241 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3301 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     3361 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3421 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3481 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3541 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3601 ccttcttgac gagttcttct gaattgaaaa aggaagagta tgagtattca acatttccgt
     3661 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3721 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3781 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3841 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     3901 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3961 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4021 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4081 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4141 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4201 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4261 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4321 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4381 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     4441 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     4501 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     4561 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4621 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4681 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4741 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4801 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4861 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4921 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4981 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5041 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5101 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5161 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5221 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5281 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5341 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5401 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
