LOCUS       sequence-225499-        5116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon optimised transposase sequence in an optimal vector for in 
            vitro transcription. Contains the T3 promoter and synthetic polyA 
            tract..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5116)
  AUTHORS   Hall TE, Martel N, Ariotti N, Xiong Z, Lo HP, Ferguson C, Rae J, Lim
            YW, Parton RG
  TITLE     In vivo cell biological screening identifies an endocytic capture 
            mechanism for T-tubule formation.
  JOURNAL   Nat Commun. 2020 Jul 24;11(1):3711. doi: 10.1038/s41467-020-17486-w.
  PUBMED    32709891
REFERENCE   2  (bases 1 to 5116)
  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: "Nat 
            Commun."; date: "2020-07-24"; volume: "11(1):3711. doi"; pages: " 
            10.1038/s41467-020-17486-w"
COMMENT     Sequence Label: pT3TS_Transposase
FEATURES             Location/Qualifiers
     source          1..5116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           44..86
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      102..111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             108..2057
                     /codon_start=1
                     /product="transposase of the Tol2 transposon from the 
                     medaka fish Oryzias latipes (Kawakami, 2007)"
                     /label=Tol2 transposase
                     /translation="MEEVCDSSAAASSTVQNQPQDQEHPWPYLREFFSLSGVNKDSFKM
                     KCVLCLPLNKEISAFKSSPSNLRKHIERMHPNYLKNYSKLTAQKRKIGTSTHASSSKQL
                     KVDSVFPVKHVSPVTVNKAILRYIIQGLHPFSTVDLPSFKELISTLQPGISVITRPTLR
                     SKIAEAALIMKQKVTAAMSEVEWIATTTDCWTARRKSFIGVTAHWINPGSLERHSAALA
                     CKRLMGSHTFEVLASAMNDIHSEYEIRDKVVCTTTDSGSNFMKAFRVFGVENNDIETEA
                     RRCESDDTDSEGCGEGSDGVEFQDASRVLDQDDGFEFQLPKHQKCACHLLNLVSSVDAQ
                     KALSNEHYKKLYRSVFGKCQALWNKSSRSALAAEAVESESRLQLLRPNQTRWNSTFMAV
                     DRILQICKEAGEGALRNICTSLEVPMFNPAEMLFLTEWANTMRPVAKVLDILQAETNTQ
                     LGWLLPSVHQLSLKLQRLHHSLRYCDPLVDALQQGIQTRFKHMFEDPEIIAAAILLPKF
                     RTSWTNDETIIKRGMDYIRVHLEPLDHKKELANSSSDDEDFFASLKPTTHEASKELDGY
                     LACVSDTRESLLTFPAICSLSIKTNTPLPASAACERLFSTAGLLFSPKRARLDTNNFEN
                     QLLLKLNLRFYNFE"
     3'UTR           2093..2218
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2304..2323)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2305..2323)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2330..2347)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2330..2346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2339..2361)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2488..2943)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2625..2646)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2837..2856
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2970..3074
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3075..3143,3144..3935)
                     /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(3293..3312)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4106..4694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4595..4614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4848..4865
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4982..5003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5018..5023,5024..5041,5042..5048)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5056..5072
                     /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     5061..5083
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5080..5096
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5080..5096
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct ggagctaagc ttgcttgttc tttttgcaga
       61 agctcagaat aaacgctcaa ctttggcaga tctaactcga cgccaccatg gaagaggtgt
      121 gcgactcttc cgccgcagct tcatctaccg tccagaacca gcctcaggac caagagcacc
      181 cctggccata tctgagagag ttcttctccc tgagcggagt gaataaggac tcatttaaaa
      241 tgaagtgtgt gctgtgtctt ccactcaaca aagagattag cgcgtttaaa tcctcaccct
      301 cgaacctgag gaagcacatt gagcgtatgc accctaacta cctgaaaaat tactccaagc
      361 tgaccgccca gaagagaaag ataggcacta gtactcacgc atccagcagt aagcagttga
      421 aagtagactc agtgttccct gttaagcatg tgtcccctgt aacagttaac aaggctattc
      481 tccggtacat tatccaaggt ctgcacccgt tcagcaccgt cgacctaccc tccttcaagg
      541 agctgattag tacgcttcag cctggaatta gtgtgattac tcgccctact ctgagaagca
      601 aaatcgcaga agcggcgctg atcatgaaac agaaagtgac tgctgcaatg agcgaggtgg
      661 aatggatcgc gacgacaacc gactgttgga cagcacgtag gaagtccttt ataggcgtca
      721 ctgcccactg gattaacccc ggttcactgg agcgacactc agcagccctg gcctgcaaac
      781 gccttatggg atctcacact tttgaggtgt tagcgtctgc tatgaacgac atccactcgg
      841 aatacgaaat tagggacaag gtggtctgta ccacaacaga ctctggatct aactttatga
      901 aagcgtttcg cgtttttggc gtggagaaca atgatatcga aacggaggcc agacgctgtg
      961 agtcagacga cactgacagc gaaggctgcg gcgaaggttc agacggcgtc gaatttcagg
     1021 acgcgagtcg cgttctcgac caggatgatg gatttgaatt tcagttacca aagcaccaga
     1081 agtgtgcctg tcatcttcta aacctcgtct cttcggtcga cgcacaaaaa gctctgtcca
     1141 acgagcatta caagaagctc taccggtctg tcttcggcaa gtgtcaggcc ctttggaaca
     1201 agtcctccag atccgctctc gccgcagagg cggtagagag tgagtccaga cttcagctgc
     1261 tgagacctaa tcagacacgc tggaacagca ctttcatggc agtggaccga atcttacaaa
     1321 tctgtaaaga agccggtgag ggcgctctcc gcaacatctg tacatcactg gaagtcccta
     1381 tgtttaaccc agctgagatg ctcttcttga cagagtgggc taacactatg cggccagtcg
     1441 ctaaagtcct cgacatcctt caggccgaaa ccaatacaca gttaggctgg ctccttccaa
     1501 gcgtgcacca actcagtctg aagctccaaa gactgcatca tagtctgcgg tactgcgatc
     1561 ccctcgtgga tgctctgcag cagggaatcc agactcggtt caaacacatg ttcgaagacc
     1621 ctgagattat cgctgcagcc atccttctcc caaaatttcg aacctcgtgg acaaacgatg
     1681 agaccattat taagcgcggc atggattata tccgcgtcca ccttgaaccc ctggaccaca
     1741 aaaaagagct ggctaattca agtagtgacg atgaggattt ctttgccagc ttgaagccga
     1801 cgacacacga ggcctcaaag gagctggacg gttacctggc ctgcgtcagc gatacaaggg
     1861 agtcactgtt gaccttccct gcaatctgtt cgctgtcgat caagacaaac actcccttgc
     1921 ctgctagtgc agcgtgtgag agactcttta gcacagcagg actcctgttc agccctaaga
     1981 gagcaaggct tgataccaac aactttgaga accagttgct tctgaaactg aatttacgat
     2041 tttacaactt cgagtgagtc gacggatcct cgagcggatc tggttaccac taaaccagcc
     2101 tcaagaacac ccgaatggag tctctaagct acataatacc aacttacact ttacaaaatg
     2161 ttgtccccca aaatgtagcc attcgtatct gctcctaawa aaaaaaaagt ttcttcacat
     2221 tctaaaaaaa aaaaaaaaaa aaaaaaaaaa aacccccccc cccccctgca gggccaagtc
     2281 ggcctctaga ggtacccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg
     2341 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     2401 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     2461 agttgcgcag cctgaatggc gaatggcaaa ttgtaagcgt taatattttg ttaaaattcg
     2521 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     2581 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     2641 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2701 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     2761 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2821 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     2881 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2941 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3001 tacattcaaa tatgtatctg ctcatgagac aataaccctg ataaatgctt caataatatt
     3061 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3121 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3181 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3241 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3301 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3361 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3421 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3481 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3541 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3601 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3661 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3721 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3781 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3841 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3901 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3961 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4021 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4081 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4141 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4201 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4261 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4321 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4381 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4441 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4501 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4561 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4621 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4681 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4741 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4801 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4861 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4921 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     4981 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5041 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5101 attacgccaa gctcga
//
