LOCUS       sequence-483452-        5534 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 5534)
  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 5534)
  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..5534
                     /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"
     promoter        354..372
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1483..1530
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1568..1586)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1593..1610)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1593..1609)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1602..1624)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            1851..1960
                     /note="incP origin of transfer"
     rep_origin      2150..2578
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2237..2256)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2447..2468
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2922..3716
                     /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(2976..2995)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3586..3605
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3734..3802,3803..4594)
                     /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(3952..3971)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4765..5353
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5254..5273
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5507..5524
                     /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 gcatctagaa ttgtaatacg
      361 actcactata gggcgaattg ggcagtgaga gctggagccc ggagatgggg actgagacaa
      421 ggccccaggc cctacggggc gcagcaggtg cgagaactcc acaatgctcg aaagagtgat
      481 gggggaaccc caagtgccct tgtattgcaa gggcttttcc ctggtgtaag gagccagggg
      541 aataagggct gggtaagact ggtgccagcc gccgcggtaa cgccagcagc ttaagtggtc
      601 gccgttttta ttgggcctaa agcgttcgta gccgggtgga taagtctcta ggtaaatctt
      661 gcgtccaagc gtaaggcttc tggagatact gtctatcttg aggccgggag aggttggagg
      721 tactcccggg gtaggggtaa aatcctgtaa tcctgggggg accaccggtg gcgaaggcgt
      781 ccaactggaa cgggcctgac ggtgagggac gaaacccagg ggagcgagcc ggattagata
      841 cccgggtagt cctgggcgta aattgatgcg gactatgtgt gggggtatcc acgaagataa
      901 cttcttgtgc cgaaggaagc agtaagtccg ccggctgggg agtacgcccg caaggcctga
      961 aacttaaagg aattggcggg ggagcaccac aaagggtgga gcctgcggtt aaattggatt
     1021 caacgccgga aaactcaccg ggggcgacgg cagaatgacg gccaagctga tgactttgct
     1081 tgacgagccg agaggaggtg catggccgcc gtcagttcgt accgtgaggt gtcctcttaa
     1141 gtgaggtaac gaacgagacc cccgccctca gttaccagca aattcctatg gaatgatggg
     1201 gactctgagg ggaccgctgc tgctaaagca gaggaaggag ggggcaacgg taggtcagta
     1261 tgccccgaat cccccgggct acacgcggcc tacaatggtt acgacaatgg gttccaacac
     1321 cgaaaggtga aggtaatccc ctaaacgtaa tctcagtccg gattgtgggc tgtaactcgc
     1381 cctcatgaag ctggaatccc tagtaatcgc gtgtcactat cgcgcggtga tacgtcctgc
     1441 tccttgcaca caccgcccgt catatcacct gagtagggct tgctagcata accccttggg
     1501 gcctctaaac gggtcttgag gggttttttg tgcatgctcg agcggccgcc agtgtgatgg
     1561 aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac
     1621 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     1681 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     1741 ggcgaatgga cgcgctggaa ggcagtacac cttgataggt gggctgccct tcctggttgg
     1801 cttggtttca tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc
     1861 tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga
     1921 acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca
     1981 ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg
     2041 atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg aaaagcgctg
     2101 cttccctgct gttttgtgga atatctaccg actggaaaca ggcaaatgca ggaaccctgt
     2161 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2221 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2281 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2341 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2401 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2461 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2521 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2581 aacaaaattc agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa gccagtccgc
     2641 agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg
     2701 caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg
     2761 cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg
     2821 ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg
     2881 gatcaagatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     2941 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3001 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3061 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3121 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3181 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatcccacc
     3241 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3301 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3361 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3421 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3481 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3541 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3601 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3661 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgaattg
     3721 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3781 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3841 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3901 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3961 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4021 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4081 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4141 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4201 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4261 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4321 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4381 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4441 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4501 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4561 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4621 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4681 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4741 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4801 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4861 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4921 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4981 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5041 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5101 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5161 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5221 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5281 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5341 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5401 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5461 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5521 cgaggaagcg gaag
//
