LOCUS       sequence-491166-        5903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (Culexarchaeum yellowstonense LCB-24-027) 16S rRNA 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5903)
  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 5903)
  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: Culex-027
FEATURES             Location/Qualifiers
     source          1..5903
                     /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(1936..1955)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1937..1955)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1962..1979)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1962..1978)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1971..1993)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2220..2329
                     /note="incP origin of transfer"
     rep_origin      2519..2947
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2606..2625)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2816..2837
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3291..4085
                     /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(3345..3364)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3955..3974
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4103..4171,4172..4963)
                     /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(4321..4340)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5134..5722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5623..5642
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5876..5893
                     /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 atgctagagg aggtgatccg gccgcaggtt cccctacggc caccttgtta cgacttctcc
      481 ctcctcatcg agcttagatt cgccctgacc ctttgcgagc cagagctcat ctaagcccga
      541 ctcggatgga gcgacgggcg gtgtgtgcaa ggagcaggga cgtattcacc gcgcgttggt
      601 gacgcgcggt tactagggat tccatgttca cgagggcgag ttgcagccct cgatcccaac
      661 tgaggtgggg tttatgggat taccttcccc tttcggggtc ggaacccatt gtccccacca
      721 ttgcagcccg cgtgtggccc gggggtttcg gggcatacgg acctgccgtg gccccctcct
      781 tcctccgcct tagcggcggc agtccgttta gtgtgcctta gcccttacta gggctaagta
      841 gcaactaaac gtgggggtct tgttcgttgc cggacttaac cggacacctc acggcacgaa
      901 ctggcgacgg ccatgcacct cctctcagct cgtctggcaa ggtcgtcagc ctggccttca
      961 tcctgctgtc gcccccggta aggttctcgg ggttgactcc aattaaaccg caggcttcac
     1021 cccttgtggt gctcccccgc caattccttt aagtttcagc cttgcggccg tactccccag
     1081 gcggcgggct taacggcttc cctgcggcac taggtggaca caaggtccac ctaacaccta
     1141 gcccgcatcg tttacagccg ggactacggg ggtatctaat ccccttcgct cccccggctt
     1201 tcgtccctca ccgtcgggcg cgttccagct gagcgccttc gccactggtg gtcctcctgg
     1261 gattatagga tttcgcccct accccaggag taccctcagc ctctcccgcc ccctagcccg
     1321 gcagtatttt cagccttccc agggttgagc cctgggcttt agctgaaaac ttaccgggcc
     1381 ggctacggac gctttaggcc caataaacgc cccgaccact cgcggagctg gtattaccgc
     1441 ggcggctgac accagacttg ccccccgctt attgcctccg gctttttaca ccggagaaaa
     1501 gccaccctca gcgggtggca ctgggggtga ccacatcgtg ctttcgcaca ttgtgtaggt
     1561 ttcgcgcctg ctgcgccccg tagggcctgg gcccttgtct cagtgcccat ctccgggctc
     1621 ccgctttcac ggcccgtacc cgttataggc ttggtgggcc gttaccccac caactacctg
     1681 atgggacgtg gccccatcct agggcgatgc caagaggcat gttcttggca tcctttcgga
     1741 gaagaggcat tccagcactc ttctcctatt ggggattatc cccagtttcc cggggttatc
     1801 cccatcccta gggtaggtta gccacgtttt actgagccgt acgccgcacc ccaacgtggc
     1861 ctggggtgct gactcgcatg gcttagtccc acccccatag cagtcgggtc cggcaggatc
     1921 aaccggggta attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa
     1981 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     2041 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2101 agcctgaatg gcgaatggac gcgctggaag gcagtacacc ttgataggtg ggctgccctt
     2161 cctggttggc ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc
     2221 cggccagcct cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt
     2281 gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg
     2341 ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa
     2401 aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga
     2461 aaagcgctgc ttccctgctg ttttgtggaa tatctaccga ctggaaacag gcaaatgcag
     2521 gaaccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2581 acacttgcta gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2641 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2701 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2761 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2821 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2881 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2941 gcgaatttta acaaaattca gggcgcaagg gctgctaaag gaagcggaac acgtagaaag
     3001 ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa
     3061 gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc
     3121 tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg
     3181 gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat
     3241 ggcgcagggg atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3301 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3361 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3421 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3481 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3541 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3601 catcccacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3661 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3721 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3781 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3841 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3901 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3961 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4021 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4081 tctgaattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4141 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4201 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4261 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4321 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4381 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4441 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4501 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4561 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4621 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4681 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4741 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4801 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4861 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4921 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4981 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5041 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5101 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5161 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5221 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5281 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5341 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5401 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5461 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5521 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5581 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5641 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5701 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5761 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5821 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5881 gtcagtgagc gaggaagcgg aag
//
