LOCUS       sequence-491205-        5504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (Uncultured archaeon clone Z7ME17) 16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5504)
  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 5504)
  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: Z7ME17
FEATURES             Location/Qualifiers
     source          1..5504
                     /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(1537..1556)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1538..1556)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1563..1580)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1563..1579)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1572..1594)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            1821..1930
                     /note="incP origin of transfer"
     rep_origin      2120..2548
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2207..2226)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2417..2438
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2892..3686
                     /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(2946..2965)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3556..3575
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3704..3772,3773..4564)
                     /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(3922..3941)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4735..5323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5224..5243
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5477..5494
                     /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 atgctagcaa gccctactca ggtgatatga cgggcggtgt gtgcaaggag caggacgtat
      481 caccgcgcga tagtgacacg cgattactag ggattccagc ttcatgaggg cgagttacag
      541 cccacaatcc ggactgagat tacgtttagg ggattacctt cacctttcgg tgttggaacc
      601 cattgtcgta accattgtag gccgcgtgta gcccggggga ttcggggcat actgacctac
      661 cgttgccccc tccttcctct gctttagcag cagcggtccc ctcagagtcc ccatcattcc
      721 ataggaattt gctggtaact gagggcgggg gtctcgttcg ttacctcact taagaggaca
      781 cctcacggta cgaactgacg gcggccatgc acctcctctc ggctcgtcaa gcaaagtcat
      841 cagcttggcc gtcattctgc cgtcgccccc ggtgagtttt ccggcgttga atccaattta
      901 accgcaggct ccaccctttg tggtgctccc ccgccaattc ctttaagttt caggccttgc
      961 gggcgtactc cccagccggc ggacttactg cttccttcgg cacaagaagt tatcttcgtg
     1021 gataccccca cacatagtcc gcatcaattt acgcccagga ctacccgggt atctaatccg
     1081 gctcgctccc ctgggtttcg tccctcaccg tcaggcccgt tccagttgga cgccttcgcc
     1141 accggtggtc cccccaggat tacaggattt tacccctacc ccgggagtac ctccaacctc
     1201 tcccggcctc aagatagaca gtatctccag aagccttacg cttggacgca agatttacct
     1261 agagacttat ccacccggct acgaacgctt taggcccaat aaaaacggcg accacttaag
     1321 ctgctggcgt taccgcggcg gctggcacca gtcttaccca gcccttattc ccctggctcc
     1381 ttacaccagg gaaaagccct tgcaatacaa gggcacttgg ggttccccca tcactctttc
     1441 gagcattgtg gagttctcgc acctgctgcg ccccgtaggg cctggggcct tgtctcagtc
     1501 cccatctccg ggctccagct ctcactgccc aattcgccct atagtgagtc gtattacaat
     1561 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     1621 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     1681 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgctggaa ggcagtacac
     1741 cttgataggt gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc
     1801 atctgttacg ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc
     1861 caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc
     1921 tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca
     1981 aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc
     2041 gaagcagggt tatgcagcgg aaaagcgctg cttccctgct gttttgtgga atatctaccg
     2101 actggaaaca ggcaaatgca ggaaccctgt agcggcgcat taagcgcggc gggtgtggtg
     2161 gttacgcgca gcgtgaccgc tacacttgct agcgccctag cgcccgctcc tttcgctttc
     2221 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2281 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2341 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2401 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2461 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2521 ctgatttaac aaaaatttaa cgcgaatttt aacaaaattc agggcgcaag ggctgctaaa
     2581 ggaagcggaa cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg atgaatgtca
     2641 gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca
     2701 gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat
     2761 tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt
     2821 tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgag
     2881 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2941 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3001 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3061 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3121 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3181 tgccggggca ggatctcctg tcatcccacc ttgctcctgc cgagaaagta tccatcatgg
     3241 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3301 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3361 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     3421 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3481 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     3541 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     3601 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     3661 atcgccttct tgacgagttc ttctgaattg aaaaaggaag agtatgagta ttcaacattt
     3721 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3781 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3841 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3901 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3961 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4021 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4081 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4141 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4201 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4261 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4321 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4381 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4441 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4501 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4561 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4621 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4681 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4741 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4801 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4861 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4921 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4981 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5041 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5101 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5161 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5221 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5281 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5341 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5401 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5461 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
