LOCUS       sequence-491196-        5899 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 5899)
  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 5899)
  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..5899
                     /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(1932..1951)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1933..1951)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1958..1975)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1958..1974)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1967..1989)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2216..2325
                     /note="incP origin of transfer"
     rep_origin      2515..2943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2602..2621)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2812..2833
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3287..4081
                     /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(3341..3360)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3951..3970
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4099..4167,4168..4959)
                     /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(4317..4336)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5130..5718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5619..5638
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5872..5889
                     /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 cccctgcggc caccttgtta cgacttctcc
      481 ccccttgcca aacatgggtt cgacccaaac aacaggtctg ggcctcgccc aagcttgact
      541 cgggtggagc gacgggcggt gtgtgcaagg agcagggacg tattcaccgc acgatggtga
      601 catgcggtta ctagggattc catgttcatg agggcgggtt acagccctca atcccaacta
      661 cggcgaggtt taagggatta ccttcccctt tcggggtcgg aacccgctgt cctcaccatt
      721 gcagcccgcg tgtggcccgg gagattcggg gcatgctgac ctgccgtagc cccctccctc
      781 ctccagttta acactggcag tctcctccgt gtgcgcctac ctccgtagaa gcaggctagc
      841 aacagaggat gggggtctcg ttcgttgccg gacttaaccg gacacctcac ggcacgaact
      901 ggcgacggcc atgcacctcc tctcagccca tctagcaaag tcgtcagctt ggccttcatc
      961 ttgctgtcgc tcccggtaag attcccggcg ttgactccaa ttaaaccgca ggcttcaccc
     1021 cttgtggtgc tcccccgcca attcctttaa gtttcagcct tgcagccgta cttcccaggc
     1081 ggcgggctta tcgtcttcac tgcggcaccg ggttggcacg aagccaaccc aacacctagc
     1141 ccgcatcgtt tacagccagg actacggggg tatctaatcc cctttgctcc cctggctttc
     1201 gtccctcacc gtcgggcgcg ttccagttga gcgccttcgc cactggtggt ccttcgcgga
     1261 ttataggatt tcgcccctac ccgcgaagta ccctcaacct ctcccgcccc ctagcctaac
     1321 agtatctcca gcaatctcaa ggttgagcct tgaggtttaa ccagagactt gataggccgg
     1381 ctacggacgc tttaggccca ataatcgtcc cgaccactcg cggagctggt attaccgcgg
     1441 cggctgacac caggcttgcc ccccgcttat tcccttagct tactacacta aggaaaagct
     1501 aacctcagcg gctagcactc ggggtggccc tgtcgcgctt tcgcgcattg cagaggtttc
     1561 gcgcctgctg cgccccgtag ggcctgggcc cttgtctcag tgcccatctc cgggctcccc
     1621 ctctcagggc ccgtacccgt tataggcttg gtgggccgtt accccaccaa ctacctgata
     1681 gggtgcagtc tcatcctcag gcggcacact gcgcatgagc aatatgccgt ttgaataatg
     1741 aaccattcca ggatccatta tctatcggat attagcctca gtttcccgag gttatctccg
     1801 tcctgagggt agattgacca catgttactg agccgtacgc cgccctccat cgtggctagg
     1861 agagctaact cgcatggctt agcctcaccc caatagcagt cgggtccggc aggatcaacc
     1921 gggataattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     1981 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     2041 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     2101 tgaatggcga atggacgcgc tggaaggcag tacaccttga taggtgggct gcccttcctg
     2161 gttggcttgg tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc
     2221 cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag
     2281 aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg
     2341 atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag
     2401 gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaag
     2461 cgctgcttcc ctgctgtttt gtggaatatc taccgactgg aaacaggcaa atgcaggaac
     2521 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2581 ttgctagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2641 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2701 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2761 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2821 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2881 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2941 attttaacaa aattcagggc gcaagggctg ctaaaggaag cggaacacgt agaaagccag
     3001 tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct ggacaaggga
     3061 aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc gatagctaga
     3121 ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctggtaa
     3181 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
     3241 caggggatca agatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3301 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3361 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3421 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3481 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3541 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3601 ccaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3661 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3721 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3781 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3841 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3901 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3961 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4021 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4081 aattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4141 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4201 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4261 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4321 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4381 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4441 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4501 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4561 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4621 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4681 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4741 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4801 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4861 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4921 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4981 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5041 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5101 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5161 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5221 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5281 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5341 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5401 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5461 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5521 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5581 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5641 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5701 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5761 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5821 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5881 gtgagcgagg aagcggaag
//
