LOCUS       sequence-483441-        5929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (Methanosuratincola verstraetei LCB70) 16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5929)
  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 5929)
  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: LCB70
FEATURES             Location/Qualifiers
     source          1..5929
                     /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      1878..1925
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1963..1981)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1988..2005)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1988..2004)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1997..2019)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2246..2355
                     /note="incP origin of transfer"
     rep_origin      2545..2973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2632..2651)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2842..2863
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3317..4111
                     /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(3371..3390)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3981..4000
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4129..4197,4198..4989)
                     /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(4347..4366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5160..5748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5649..5668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5902..5919
                     /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 tcccggttga tcctgccgga cccgactgct attggggtga
      421 ggctaagcca tgcgagttag ctctcctagc cacgatggag ggcggcgtac ggctcagtaa
      481 catgtggtca atctaccctc aggacggaga taacctcggg aaactgaggc taatatccga
      541 tagataatgg atcctggaat ggttcattat tcaaacggca tattgctcat gcgcagtgtg
      601 ccgcctgagg atgagactgc accctatcag gtagttggtg gggtaacggc ccaccaagcc
      661 tataacgggt acgggccctg agagggggag cccggagatg ggcactgaga caagggccca
      721 ggccctacgg ggcgcagcag gcgcgaaacc tctgcaatgc gcgaaagcgc gacagggcca
      781 ccccgagtgc tagccgctga ggttagcttt tccttagtgt agtaagctaa gggaataagc
      841 ggggggcaag cctggtgtca gccgccgcgg taataccagc tccgcgagtg gtcgggacga
      901 ttattgggcc taaagcgtcc gtagccggcc tatcaagtct ctggttaaac ctcaaggctc
      961 aaccttgaga ttgctggaga tactgttagg ctagggggcg ggagaggttg agggtacttc
     1021 gcgggtaggg gcgaaatcct ataatccgcg aaggaccacc agtggcgaag gcgctcaact
     1081 ggaacgcgcc cgacggtgag ggacgaaagc caggggagca aaggggatta gatacccccg
     1141 tagtcctggc tgtaaacgat gcgggctagg tgttgggttg gcttcgtgcc aacccggtgc
     1201 cgcagtgaag acgataagcc cgccgcctgg gaagtacggc tgcaaggctg aaacttaaag
     1261 gaattggcgg gggagcacca caaggggtga agcctgcggt ttaattggag tcaacgccgg
     1321 gaatcttacc gggagcgaca gcaagatgaa ggccaagctg acgactttgc tagatgggct
     1381 gagaggaggt gcatggccgt cgccagttcg tgccgtgagg tgtccggtta agtccggcaa
     1441 cgaacgagac ccccatcctc tgttgctagc ctgcttctac ggaggtaggc gcacacggag
     1501 gagactgcca gtgttaaact ggaggaggga gggggctacg gcaggtcagc atgccccgaa
     1561 tctcccgggc cacacgcggg ctgcaatggt gaggacagcg ggttccgacc ccgaaagggg
     1621 aaggtaatcc cttaaacctc gccgtagttg ggattgaggg ctgtaacccg ccctcatgaa
     1681 catggaatcc ctagtaaccg catgtcacca tcgtgcggtg aatacgtccc tgctccttgc
     1741 acacaccgcc cgtcgctcca cccgagtcaa gcttgggcga ggcccagacc tgttgtttgg
     1801 gtcgaaccca tgtttggcaa ggggggagaa gtcgtaacaa ggtggccgca ggggaacctg
     1861 cggccggatc acctcctcta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1921 ttttgtgcat gctcgagcgg ccgccagtgt gatggaattc gccctatagt gagtcgtatt
     1981 acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     2041 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     2101 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc tggaaggcag
     2161 tacaccttga taggtgggct gcccttcctg gttggcttgg tttcatcagc catccgcttg
     2221 ccctcatctg ttacgccggc ggtagccggc cagcctcgca gagcaggatt cccgttgagc
     2281 accgccaggt gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact
     2341 tcacctatcc tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt
     2401 tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg
     2461 accccgaagc agggttatgc agcggaaaag cgctgcttcc ctgctgtttt gtggaatatc
     2521 taccgactgg aaacaggcaa atgcaggaac cctgtagcgg cgcattaagc gcggcgggtg
     2581 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2641 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2701 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2761 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2821 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2881 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2941 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aattcagggc gcaagggctg
     3001 ctaaaggaag cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa
     3061 tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc
     3121 ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc
     3181 ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat
     3241 ggctttcttg ccgccaagga tctgatggcg caggggatca agatctgatc aagagacagg
     3301 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3361 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3421 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3481 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3541 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3601 cgaagtgccg gggcaggatc tcctgtcatc ccaccttgct cctgccgaga aagtatccat
     3661 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3721 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3781 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3841 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3901 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3961 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4021 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4081 cttctatcgc cttcttgacg agttcttctg aattgaaaaa ggaagagtat gagtattcaa
     4141 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4201 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4261 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4321 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4381 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4441 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4501 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4561 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4621 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4681 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4741 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4801 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4861 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4921 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4981 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5041 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5101 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5161 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5221 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5281 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5341 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5401 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5461 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5521 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5581 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5641 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5701 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5761 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5821 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5881 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
