LOCUS       40924_42304        4247 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pT3TS-Cre, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4247)
  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  JOURNAL   Unpublished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 4247)
  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 4247)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4247)
  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            "Unpublished"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..4247
                     /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             99..1127
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     3'UTR           1163..1287
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus
                     laevis"
     promoter        complement(1406..1424)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1431..1447)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(1588..2043)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2070..2174
                     /label=AmpR promoter
     CDS             2175..3032
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3206..3794
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4082..4103
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4118..4148
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4156..4172
                     /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     4180..4196
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4217..4235
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgcttgttc tttttgcaga
       61 agctcagaat aaacgctcaa ctttggcaga tctacaagat gtccaattta ctgaccgtac
      121 accaaaattt gcctgcatta ccggtcgatg caacgagtga tgaggttcgc aagaacctga
      181 tggacatgtt cagggatcgc caggcgtttt ctgagcatac ctggaaaatg cttctgtccg
      241 tttgccggtc gtgggcggca tggtgcaagt tgaataaccg gaaatggttt cccgcagaac
      301 ctgaagatgt tcgcgattat cttctatatc ttcaggcgcg cggtctggca gtaaaaacta
      361 tccagcaaca tttgggccag ctaaacatgc ttcatcgtcg gtccgggctg ccacgaccaa
      421 gtgacagcaa tgctgtttca ctggttatgc ggcggatccg aaaagaaaac gttgatgccg
      481 gtgaacgtgc aaaacaggct ctagcgttcg aacgcactga tttcgaccag gttcgttcac
      541 tcatggaaaa tagcgatcgc tgccaggata tacgtaatct ggcatttctg gggattgctt
      601 ataacaccct gttacgtata gccgaaattg ccaggatcag ggttaaagat atctcacgta
      661 ctgacggtgg gagaatgtta atccatattg gcagaacgaa aacgctggtt agcaccgcag
      721 gtgtagagaa ggcacttagc ctgggggtaa ctaaactggt cgagcgatgg atttccgtct
      781 ctggtgtagc tgatgatccg aataactacc tgttttgccg ggtcagaaaa aatggtgttg
      841 ccgcgccatc tgccaccagc cagctatcaa ctcgcgccct ggaagggatt tttgaagcaa
      901 ctcatcgatt gatttacggc gctaaggatg actctggtca gagatacctg gcctggtctg
      961 gacacagtgc ccgtgtcgga gccgcgcgag atatggcccg cgctggagtt tcaataccgg
     1021 agatcatgca agctggtggc tggaccaatg taaatattgt catgaactat atccgtaacc
     1081 tggatagtga aacaggggca atggtgcgcc tgctggaaga tggcgattaa actagtgact
     1141 gactaggatc tggttaccac taaaccagcc tcaagaacac ccgaatggag tctctaagct
     1201 acataatacc aacttacact tacaaaatgt tgtcccccaa aatgtagcca ttcgtatctg
     1261 ctcctaataa aaagaaagtt tcttcacatt ctaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     1321 aacccccccc cccccccccc cccccccccc ccgcatgcct gcaggtcgac tctagaggat
     1381 ccccgggtac cgagctccaa ttcgccctat agtgagtcgt attacaattc actggccgtc
     1441 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     1501 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     1561 cagttgcgca gcctgaatgg cgaatggaaa ttgtaagcgt taatattttg ttaaaattcg
     1621 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     1681 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     1741 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     1801 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     1861 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     1921 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     1981 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2041 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2101 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2161 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2221 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2281 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2341 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2401 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2461 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2521 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2581 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2641 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2701 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     2761 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2821 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2881 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2941 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3001 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3061 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3121 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3181 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3241 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3301 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3361 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3421 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3481 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3541 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3601 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3661 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3721 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3781 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3841 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3901 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3961 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4021 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     4081 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     4141 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     4201 attacgccaa gctcgaaatt aaccctcact aaagggaaca aaagctg
//
