LOCUS       40924_42309        5163 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pT3TS-Tol2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5163)
  AUTHORS   Clark KJ, Balciunas D, Pagoda H-M., Ding Y, Westcot SE, Bedell VM, 
            Greenwood TM, Urban MD, Skuster KJ, Petzold AM, Ni J, Nielson A, 
            Sivasubbu S, Xu X, Hammerschmidt M, Ekker SC.
  TITLE     In vivo protein trapping produces a functional expression codex of 
            the vertebrate proteome
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5163)
  AUTHORS   Clark KJ, Balciunas D, Pagoda H-M., Ding Y, Westcot SE, Bedell VM, 
            Greenwood TM, Urban MD, Skuster KJ, Petzold AM, Ni J, Nielson A, 
            Sivasubbu S, Xu X, Hammerschmidt M, Ekker SC.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-OCT-2010) Biochemistry and Molecular Biology, Mayo 
            Clinic, 221 4th Ave. SW, Rochester, MN 55902, USA
REFERENCE   3  (bases 1 to 5163)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5163)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-OCT-2010) Biochemistry and Molecular Biology, Mayo Clinic, 221 
            4th Ave. SW, Rochester, MN 55902, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..34
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     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"
     CDS             97..2043
                     /codon_start=1
                     /label=Tol2
                     /note="Transposase encoded from Tol2 transposon, a member
                     of hAT transposable element family found in medaka fish 
                     genome"
                     /translation="MEEVCDSSAAASSTVQNQPQDQEHPWPYLREFFSLSGVNKDSFKM
                     KCVLCLPLNKEISAFKSSPSNLRKHIERMHPNYLKNYSKLTAQKRKIGTSTHASSSKQL
                     KVDSVFPVKHVSPVTVNKAILRYIIQGLHPFSTVDLPSFKELISTLQPGISVITRPTLR
                     SKIAEAALIMKQKVTAAMSEVEWIATTTDCWTARRKSFIGVTAHWINPGSLERHSAALA
                     CKRLMGSHTFEVLASAMNDIHSEYEIRDKVVCTTTDSGSNFMKAFRVFGVENNDIETEA
                     RRCESDDTDSEGCGEGSDGVEFQDASRVLDQDDGFEFQLPKHQKCACHLLNLVSSVDAQ
                     KALSNEHYKKLYRSVFGKCQALWNKSSRSALAAEAVESESRLQLLRPNQTRWNSTFMAV
                     DRILQICKEAGEGALRNICTSLEVPMFNPAEMLFLTEWANTMRPVAKVLDILQAETNTQ
                     LGWLLPSVHQLSLKLQRLHHSLRYCDPLVDALQQGIQTRFKHMFEDPEIIAAAILLPKF
                     RTSWTNDETIIKRGMDYIRVHLEPLDHKKELANSSSDDEDFFASLKPTTHEASKELDGY
                     LACVSDTRESLLTFPAICSLSIKTNTPLPASAACERLFSTAGLLFSPKRARLDTNNFEN
                     QLLLKLNLRFYNFE"
     3'UTR           2079..2203
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus
                     laevis"
     promoter        complement(2322..2340)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2347..2363)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2504..2959)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2986..3090
                     /label=AmpR promoter
     CDS             3091..3948
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4122..4710
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4998..5019
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5034..5064
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5072..5088
                     /label=lac operator
                     /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     5096..5112
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5133..5151
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgcttgttc tttttgcaga
       61 agctcagaat aaacgctcaa ctttggcaga tccaccatgg aggaagtatg tgattcatca
      121 gcagctgcga gcagcacagt ccaaaatcag ccacaggatc aagagcaccc gtggccgtat
      181 cttcgcgaat tcttttcttt aagtggtgta aataaagatt cattcaagat gaaatgtgtc
      241 ctctgtctcc cgcttaataa agaaatatcg gccttcaaaa gttcgccatc aaacctaagg
      301 aagcatattg agagaatgca cccaaattac ctcaaaaact actctaaatt gacagcacag
      361 aagagaaaga tcgggacctc cacccatgct tccagcagta agcaactgaa agttgactca
      421 gttttcccag tcaaacatgt gtctccagtc actgtgaaca aagctatatt aaggtacatc
      481 attcaaggac ttcatccttt cagcactgtt gatctgccat catttaaaga gctgattagt
      541 acactgcagc ctggcatttc tgtcattaca aggcctactt tacgctccaa gatagctgaa
      601 gctgctctga tcatgaaaca gaaagtgact gctgccatga gtgaagttga atggattgca
      661 accacaacgg attgttggac tgcacgtaga aagtcattca ttggtgtaac tgctcactgg
      721 atcaaccctg gaagtcttga aagacattcc gctgcacttg cctgcaaaag attaatgggc
      781 tctcatactt ttgaggtact ggccagtgcc atgaatgata tccactcaga gtatgaaata
      841 cgtgacaagg ttgtttgcac aaccacagac agtggttcca actttatgaa ggctttcaga
      901 gtttttggtg tggaaaacaa tgatatcgag actgaggcaa gaaggtgtga aagtgatgac
      961 actgattctg aaggctgtgg tgagggaagt gatggtgtgg aattccaaga tgcctcacga
     1021 gtcctggacc aagacgatgg cttcgaattc cagctaccaa aacatcaaaa gtgtgcctgt
     1081 cacttactta acctagtctc aagcgttgat gcccaaaaag ctctctcaaa tgaacactac
     1141 aagaaactct acagatctgt ctttggcaaa tgccaagctt tatggaataa aagcagccga
     1201 tcggctctag cagctgaagc tgttgaatca gaaagccggc ttcagctttt aaggccaaac
     1261 caaacgcggt ggaattcaac ttttatggct gttgacagaa ttcttcaaat ttgcaaagaa
     1321 gcaggagaag gcgcacttcg gaatatatgc acctctcttg aggttccaat gtttaatcca
     1381 gcagaaatgc tgttcttgac agagtgggcc aacacaatgc gtccagttgc aaaagtactc
     1441 gacatcttgc aagcggaaac gaatacacag ctggggtggc tgctgcctag tgtccatcag
     1501 ttaagcttga aacttcagcg actccaccat tctctcaggt actgtgaccc acttgtggat
     1561 gccctacaac aaggaatcca aacacgattc aagcatatgt ttgaagatcc tgagatcata
     1621 gcagctgcca tccttctccc taaatttcgg acctcttgga caaatgatga aaccatcata
     1681 aaacgaggca tggactacat cagagtgcat ctggagcctt tggaccacaa gaaggaattg
     1741 gccaacagtt catctgatga tgaagatttt ttcgcttctt tgaaaccgac aacacatgaa
     1801 gccagcaaag agttggatgg atatctggcc tgtgtttcag acaccaggga gtctctgctc
     1861 acgtttcctg ctatttgcag cctctctatc aagactaata cacctcttcc cgcatcggct
     1921 gcctgtgaga ggcttttcag cactgcagga ttgcttttca gccccaaaag agctaggctt
     1981 gacactaaca attttgagaa tcagcttcta ctgaagttaa atctgaggtt ttacaacttt
     2041 gagtagacta gtgactgact aggatctggt taccactaaa ccagcctcaa gaacacccga
     2101 atggagtctc taagctacat aataccaact tacacttaca aaatgttgtc ccccaaaatg
     2161 tagccattcg tatctgctcc taataaaaag aaagtttctt cacattctaa aaaaaaaaaa
     2221 aaaaaaaaaa aaaaaaaacc cccccccccc cccccccccc ccccccccgc atgcctgcag
     2281 gtcgactcta gaggatcccc gggtaccgag ctccaattcg ccctatagtg agtcgtatta
     2341 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2401 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2461 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggaaattgt aagcgttaat
     2521 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2581 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2641 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2701 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2761 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2821 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2881 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2941 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3001 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3061 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3121 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3181 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3241 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3301 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3361 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3421 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3481 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3541 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3601 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3661 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3721 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3781 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3841 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3901 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3961 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4021 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4081 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4141 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4201 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4261 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4321 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4381 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4441 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4501 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4561 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4621 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4681 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4741 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4801 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4861 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4921 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4981 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5041 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     5101 aacagctatg accatgatta cgccaagctc gaaattaacc ctcactaaag ggaacaaaag
     5161 ctg
//
