LOCUS       sequence-491204-        5839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Archael (Thalassoarchaea mediterranii) 16S rRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5839)
  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 5839)
  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: Thalmedi
FEATURES             Location/Qualifiers
     source          1..5839
                     /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"
     primer_bind     complement(1872..1891)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1873..1891)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1898..1915)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1898..1914)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1907..1929)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     oriT            2156..2265
                     /note="incP origin of transfer"
     rep_origin      2455..2883
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2542..2561)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2752..2773
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3227..4021
                     /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(3281..3300)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3891..3910
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4039..4107,4108..4899)
                     /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(4257..4276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5070..5658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5559..5578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5812..5829
                     /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 gcaccaaaac cccctcaaga
      361 cccgtttaga ggccccaagg ggttatgcta gcgtaggagg tgatccatct gcaggttccc
      421 ctacagatac cttgttacga cttaaccctc cttgtcgaag gcaggctcga ataccccaat
      481 aagaggtagc ctcacccacc cacaactaag atggtttgac gggcggtgtg tgcaaggagc
      541 aggggcatat tcaccgcgcc ttattgagac gcgattacta cctaatccag cttcatgagg
      601 gcgagttaca gccctcaatc cgaactacga acgggtttcg ggatagcgtc attcctttcg
      661 gagtagcagc ccattgtccc gtcctttgta ttgcgcgtgt agcccatagc attcggagca
      721 tactgaccta ccgttgccct ccccttcctc taacttcgcg ctagcggtct ctctatagtc
      781 ccgtcagatc cagagatctg gtggcaaata gagatgaggg tctcgttcgt tatctgactt
      841 aacagaacgc cttacggtac gaactgacgg cggccatgca ctacctctcg aacaatctgg
      901 taagttcatt acactgacct tcatatgtcc gtcggctatg gtgagttttc cggcgttgaa
      961 tccaattaaa ccgcacaatc ctcaggttgc ggtgctcccc cgccaattcc tttaagtttc
     1021 agccttgcga ccgtacttcc caggcagtgg acttatcacc ttcgcttcga cactgaagaa
     1081 gctcggagct tctccaacat caagtccaca gcgtttacac ctaggactac ccgggtatct
     1141 aatccggttc gtgcccctag gcttcgtccc tgaccgtcgg atccgttcta gtgtggcgcc
     1201 ttcgcaacag gtggtcctct caggatgaca ggattttacc cctaccctga gagtacccca
     1261 cacctctccc ggtcccaagt ctggccgtcc tcgcagaatt caactcgttg agcgagttga
     1321 tttacccacg aatgtaccaa accggctacg gacgttttag gcccaataaa agcgactacc
     1381 acttgaggcg ccggtattac cgcggcggct ggcaccgtcc ttacccaccc cttattctac
     1441 aacctttgaa gagttatgaa aagactgggt cttaccccag ccacttggag ttccctgatc
     1501 acgattgctc gcattgtcaa gttttcgcgc ctgctgcacc ccgtagggtc tggattcgtg
     1561 tctcagaatc catctccggg ctatgtctcc caacacccgt acgcgtagat ggctagaggg
     1621 tactttacac cccctacaac ctgatacgcc gcaggcccat cctatgccgc ctgagctttg
     1681 atccactcac cgttccaggc gtagtggact atggggcatt atactcagtt tcccgaggtt
     1741 attcccctgc ataggttagg ttacccacgt gttactgagc ggtacgccga gctccgaaag
     1801 ctctggactc gcatggctta atcgaactcc aatagcggta gcctccggca ggatcaaccg
     1861 gatttaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     1921 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1981 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     2041 tgaatggcga atggacgcgc tggaaggcag tacaccttga taggtgggct gcccttcctg
     2101 gttggcttgg tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc
     2161 cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag
     2221 aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg
     2281 atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag
     2341 gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaag
     2401 cgctgcttcc ctgctgtttt gtggaatatc taccgactgg aaacaggcaa atgcaggaac
     2461 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2521 ttgctagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2581 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2641 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2701 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2761 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2821 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2881 attttaacaa aattcagggc gcaagggctg ctaaaggaag cggaacacgt agaaagccag
     2941 tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct ggacaaggga
     3001 aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc gatagctaga
     3061 ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctggtaa
     3121 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
     3181 caggggatca agatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3241 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3301 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3361 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3421 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3481 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3541 ccaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3601 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3661 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3721 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3781 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3841 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3901 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3961 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4021 aattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4081 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4141 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4201 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4261 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4321 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4381 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4441 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4501 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4561 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4621 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4681 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4741 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4801 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4861 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4921 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4981 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5041 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5101 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5161 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5221 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5281 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5341 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5401 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5461 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5521 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5581 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5641 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5701 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5761 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5821 gtgagcgagg aagcggaag
//
