LOCUS       sequence-391228-        4488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tardigrade (Hypsibius exemplaris) alpha-tubulin Tub1A1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4488)
  AUTHORS   Novotna Floriancicova K, Baltzis A, Smejkal J, Czernekova M, 
            Kaczmarek L, Maly J, Notredame C, Vinopal S
  TITLE     Phylogenetic and functional characterization of water bears 
            (Tardigrada) tubulins.
  JOURNAL   Sci Rep. 2023 Mar 30;13(1):5194. doi: 10.1038/s41598-023-31992-z.
  PUBMED    36997657
REFERENCE   2  (bases 1 to 4488)
  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: "Sci Rep."; 
            date: "2023-03-30"; volume: "13(1):5194. doi"; pages: " 
            10.1038/s41598-023-31992-z"
COMMENT     Sequence Label: pJet1.2-tHe-Tub1A1
FEATURES             Location/Qualifiers
     source          1..4488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2285..2307)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2296..2312
                     /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)."
     promoter        complement(join(2320..2326,2327..2344,2345..2350))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2365..2386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2506..2523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2677..3265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2757..2776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3436..4227,4228..4296))
                     /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     4059..4078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4297..4401)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4469..4487
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
      361 ttcagcaaga tgtgcgggga gattactgta aagcttgctt taaaactcaa gcgccttcga
      421 tcgttgttgc cgcggcgtgc ttgaagtgag agacgtcttt aaacttcctc tccctcgcct
      481 tccccctctc cctcgacaga gtccacaccg acctcctcgt agtccttctc cagggcggcc
      541 aaatcctcac gggcctcgct gaactcgcct tcctccatgc cctcaccgac gtaccagtgg
      601 acaaaggcgc gcttggcgta catcagatcg aacttgtgat ccaagcgggc ccaggcctcg
      661 gcgatggcgg tcgtgttgga gagcatacag acggcgcgct gcaccttggc caaatcaccg
      721 cccggcacaa cggtgggagg ctggtagtta attccgacct tgaagccagt cgggcaccaa
      781 tccacaaact ggatggtccg cttggttttg atcgcggcaa tggcggcgtt cacgtccttg
      841 ggcacgacgt cacctcggta gagcatgcag caggccatgt acttgccgtg gcgcggatcg
      901 cacttcacca tctggttggc cggctcgaag caagcgttgg taatctccga aacggtgagc
      961 tgttcgtggt aggccttctc ggcggaaatg atcggagcgt atgtgaccag agggaaatgg
     1021 atacgaggat aggggaccaa gttggtctgg aactcagtga gatcgacgtt gagggcaccg
     1081 tcaaaacgca gcgacgccgt gatggatgag acaatctgcc cgatcaagcg attcaagttg
     1141 gtgtaggtcg gtcgctcaat gtccagattg cggcggcaga tgtcgtagat ggcctcattg
     1201 tctaccatga aggcgcagtc agagtgctcc agggtggtgt gggtcgtcag gatagagttg
     1261 tacggctcaa ccacagcggt ggaaacctga ggcgcggggt agatggagaa ctcgagcttg
     1321 gatttcttcc catagtccac cgacagacgc tccatgagga gggaggtgaa gcccgatccg
     1381 gtgccaccgc caaagctgtg aaaaatgagg aagccctgca ggccggtgca ctggtcagcg
     1441 agcttgcgga tccggtccag caccaggtcg acgatctcct tccccacggt gtagtgaccg
     1501 cgggcgtagt tgttagccgc gtcctccttg ccagtgatca actgctcggg atggaacagc
     1561 tggcggtaag taccagttcg gatttcatca ataacagttg gctccaagtc aacaaaaacg
     1621 gcacggggca cgtgctttcc ggctccggtc tcgctgaaga aggtgttgaa gctgtcgtca
     1681 ccgccgccga tggttttgtc cgagggcatc tgcccatccg gctggatgcc atgctcgagg
     1741 cagtagagct cccagcaggc gttgccgatc tgcactccgg cctggccgac atggactgag
     1801 atgcattcac gcattttgat ggaatgtgtg gaggtgatcg cggggttgat gagacagggc
     1861 gaagaggatc tgctaaggag acgtaatctt tctagaagat ctcctacaat attctcagct
     1921 gccatggaaa atcgatgttc ttcttttatt ctctcaagat tttcaggctg tatattaaaa
     1981 cttatattaa gaactatgct aaccacctca tcaggaaccg ttgtaggtgg cgtgggtttt
     2041 cttggcaatc gactctcatg aaaactacga gctaaatatt caatatgttc ctcttgacca
     2101 actttattct gcattttttt tgaacgaggt ttagagcaag cttcaggaaa ctgagacagg
     2161 aattttatta aaaatttaaa ttttgaagaa agttcagggt taatagcatc cattttttgc
     2221 tttgcaagtt cctcagcatt cttaacaaaa gacgtctctt ttgacatgtt taaagtttaa
     2281 acctcctgtg tgaaattgtt atccgctcac aattccacac attatacgag ccggaagcat
     2341 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2401 actgccaatt gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2461 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2521 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2581 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2641 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2701 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2761 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2821 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2881 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2941 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3001 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3061 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3121 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3181 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3241 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3301 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3361 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3421 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3481 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3541 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3601 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3661 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3721 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3781 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3841 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3901 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3961 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4021 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4081 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4141 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4201 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4261 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4321 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4381 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4441 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcc
//
