LOCUS       sequence-483430-        5933 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 5933)
  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 5933)
  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..5933
                     /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      1882..1929
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1967..1985)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1992..2009)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1992..2008)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2001..2023)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2250..2359
                     /note="incP origin of transfer"
     rep_origin      2549..2977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2636..2655)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2846..2867
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3321..4115
                     /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(3375..3394)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3985..4004
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4133..4201,4202..4993)
                     /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(4351..4370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5164..5752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5653..5672
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5906..5923
                     /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 gggcgaatta ccccggttga tcctgccgga cccgactgct atgggggtgg
      421 gactaagcca tgcgagtcag caccccaggc cacgttgggg tgcggcgtac ggctcagtaa
      481 aacgtggcta acctacccta gggatgggga taaccccggg aaactgggga taatccccaa
      541 taggagaaga gtgctggaat gcctcttctc cgaaaggatg ccaagaacat gcctcttggc
      601 atcgccctag gatggggcca cgtcccatca ggtagttggt ggggtaacgg cccaccaagc
      661 ctataacggg tacgggccgt gaaagcggga gcccggagat gggcactgag acaagggccc
      721 aggccctacg gggcgcagca ggcgcgaaac ctacacaatg tgcgaaagca cgatgtggtc
      781 acccccagtg ccacccgctg agggtggctt ttctccggtg taaaaagccg gaggcaataa
      841 gcggggggca agtctggtgt cagccgccgc ggtaatacca gctccgcgag tggtcggggc
      901 gtttattggg cctaaagcgt ccgtagccgg cccggtaagt tttcagctaa agcccagggc
      961 tcaaccctgg gaaggctgaa aatactgccg ggctaggggg cgggagaggc tgagggtact
     1021 cctggggtag gggcgaaatc ctataatccc aggaggacca ccagtggcga aggcgctcag
     1081 ctggaacgcg cccgacggtg agggacgaaa gccgggggag cgaaggggat tagatacccc
     1141 cgtagtcccg gctgtaaacg atgcgggcta ggtgttaggt ggaccttgtg tccacctagt
     1201 gccgcaggga agccgttaag cccgccgcct ggggagtacg gccgcaaggc tgaaacttaa
     1261 aggaattggc gggggagcac cacaaggggt gaagcctgcg gtttaattgg agtcaacccc
     1321 gagaacctta ccgggggcga cagcaggatg aaggccaggc tgacgacctt gccagacgag
     1381 ctgagaggag gtgcatggcc gtcgccagtt cgtgccgtga ggtgtccggt taagtccggc
     1441 aacgaacaag acccccacgt ttagttgcta cttagcccta gtaagggcta aggcacacta
     1501 aacggactgc cgccgctaag gcggaggaag gagggggcca cggcaggtcc gtatgccccg
     1561 aaacccccgg gccacacgcg ggctgcaatg gtggggacaa tgggttccga ccccgaaagg
     1621 ggaaggtaat cccataaacc ccacctcagt tgggatcgag ggctgcaact cgccctcgtg
     1681 aacatggaat ccctagtaac cgcgcgtcac caacgcgcgg tgaatacgtc cctgctcctt
     1741 gcacacaccg cccgtcgctc catccgagtc gggcttagat gagctctggc tcgcaaaggg
     1801 tcagggcgaa tctaagctcg atgaggaggg agaagtcgta acaaggtggc cgtaggggaa
     1861 cctgcggccg gatcacctcc tctagcataa ccccttgggg cctctaaacg ggtcttgagg
     1921 ggttttttgt gcatgctcga gcggccgcca gtgtgatgga attcgcccta tagtgagtcg
     1981 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2041 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2101 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgctggaag
     2161 gcagtacacc ttgataggtg ggctgccctt cctggttggc ttggtttcat cagccatccg
     2221 cttgccctca tctgttacgc cggcggtagc cggccagcct cgcagagcag gattcccgtt
     2281 gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg cgggtgggcc
     2341 tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt ctacacgaac
     2401 cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat
     2461 aatgaccccg aagcagggtt atgcagcgga aaagcgctgc ttccctgctg ttttgtggaa
     2521 tatctaccga ctggaaacag gcaaatgcag gaaccctgta gcggcgcatt aagcgcggcg
     2581 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2641 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2701 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2761 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2821 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2881 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2941 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaattca gggcgcaagg
     3001 gctgctaaag gaagcggaac acgtagaaag ccagtccgca gaaacggtgc tgaccccgga
     3061 tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg
     3121 tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg
     3181 aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact
     3241 ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga
     3301 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3361 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3421 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3481 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     3541 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     3601 tgggcgaagt gccggggcag gatctcctgt catcccacct tgctcctgcc gagaaagtat
     3661 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     3721 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     3781 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     3841 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     3901 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     3961 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4021 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4081 tcgccttcta tcgccttctt gacgagttct tctgaattga aaaaggaaga gtatgagtat
     4141 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4201 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4261 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4321 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4381 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4441 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4501 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4561 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4621 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4681 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4741 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4801 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4861 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4921 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4981 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5041 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5101 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5161 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5221 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5281 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5341 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5401 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5461 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5521 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5581 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5641 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5701 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5761 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5821 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5881 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
