LOCUS       40924_42059        7289 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pT11_T11_72bp-mKate2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7289)
  AUTHORS   Li Y, Jiang Y, Chen H, Liao W, Li Z, Weiss R, Xie Z.
  TITLE     Modular construction of mammalian gene circuits using TALE 
            transcriptional repressors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7289)
  AUTHORS   Li Y, Jiang Y, Chen H, Liao W.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-SEP-2014) Bioinformatics Division/Center for Synthetic
REFERENCE   3  (bases 1 to 7289)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7289)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-SEP-2014) Bioinformatics Division/Center for Synthetic "
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..21
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     protein_bind    69..163
                     /label=5X UAS
                     /note="five tandem copies of the 'ScaI site' 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    177..196
                     /label=TALER11 bs binding site
                     /bound_moiety="TALER11 bs"
     misc_feature    203..262
                     /label=CMVmini
                     /note="CMVmini"
     protein_bind    269..288
                     /label=TALER11 bs binding site
                     /bound_moiety="TALER11 bs"
     protein_bind    305..325
                     /label=attB1
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     CDS             367..384
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             385..1080
                     /codon_start=1
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHK"
     protein_bind    complement(1123..1147)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1302..1357
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1718..1734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1742..1758
                     /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(1766..1796)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1811..1832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    2005..2841
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(2871..2889)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2910..2926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2934..2950)
                     /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)."
     promoter        complement(2958..2988)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3003..3024)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3312..3900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4074..4931)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4932..5036)
                     /label=AmpR promoter
     rep_origin      complement(5062..5517)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     5659..5675
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5685..5703
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5718..6521
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    6600..6625
                     /label=SA
                     /note="splice acceptor site"
ORIGIN
        1 caactttgta tagaaaagtt ggctccgaat ttctcgacag atctcatgtg attacgccaa
       61 gctacgggcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga gtactgtcct
      121 ccgagcggag tactgtcctc cgagcggagt tctgtcctcc gagcggagac tctagatctt
      181 tgaaagcccc gccactgaat tctaggcgtg tacggtggga ggcctatata agcagagctc
      241 gtttagtgaa ccgtcagatc gcctcgagtc tttgaaagcc ccgccactgg atccaattcg
      301 accccaagtt tgtacaaaaa agcagggatc ccggtaccct cgacacctgc tgacggagcc
      361 accatgcacc atcaccatca ccatatggtg agcgagctga ttaaggagaa catgcacatg
      421 aagctgtaca tggagggcac cgtgaacaac caccacttca agtgcacatc cgagggcgaa
      481 ggcaagccct acgagggcac ccagaccatg agaatcaagg cggtcgaggg cggccctctc
      541 cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcaaaac cttcatcaac
      601 cacacccagg gcatccccga cttctttaag cagtccttcc ccgagggctt cacatgggag
      661 agagtcacca catacgaaga tgggggcgtg ctgaccgcta cccaggacac cagcctccag
      721 gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcccatc caacggccct
      781 gtgatgcaga agaaaacact cggctgggag gcctccaccg agacactgta ccccgctgac
      841 ggcggcctgg aaggcagagc cgacatggcc ctgaagctcg tgggcggggg ccacctgatc
      901 tgcaacctta agaccacata cagatccaag aaacccgcta agaacctcaa gatgcccggc
      961 gtctactatg tggacaggag actggaaaga atcaaggagg ccgacaaaga gacatacgtc
     1021 gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact ggggcacaaa
     1081 cttaattcct agtaacgctc agtgcaggtg aattctacca ccacccagct ttcttgtaca
     1141 aagtggtacg cgtgaattca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     1201 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     1261 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     1321 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     1381 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     1441 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     1501 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     1561 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     1621 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     1681 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     1741 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1801 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1861 agtcgggaaa cctgtcgtgc cagcggatcg acagtactaa gcttggtcta atacccaatc
     1921 tctcgtctta tccagatgtt ttatacgcct taattaatag ggataacagg gtaatctcga
     1981 ggtttaaacg tcgacgcggc cgctactagg gacaggattg gtgacagaaa agccccatcc
     2041 ttaggcctcc tccttcctag tctcctgata ttgggtctaa cccccacctc ctgttaggca
     2101 gattccttat ctggtgacac acccccattt cctggagcca tctctctcct tgccagaacc
     2161 tctaaggttt gcttacgatg gagccagaga ggatcctggg agggagagct tggcaggggg
     2221 tgggagggaa gggggggatg cgtgacctgc ccggttctca gtggccaccc tgcgctaccc
     2281 tctcccagaa cctgagctgc tctgacgcgg ctgtctggtg cgtttcactg atcctggtgc
     2341 tgcagcttcc ttacacttcc caagaggaga agcagtttgg aaaaacaaaa tcagaataag
     2401 ttggtcctga gttctaactt tggctcttca cctttctagt ccccaattta tattgttcct
     2461 ccgtgcgtca gttttacctg tgagataagg ccagtagcca gccccgtcct ggcagggctg
     2521 tggtgaggag gggggtgtcc gtgtggaaaa ctccctttgt gagaatggtg cgtcctaggt
     2581 gttcaccagg tcgtggccgc ctctactccc tttctctttc tccatccttc tttccttaaa
     2641 gagtccccag tgctatctgg gacatattcc tccgcccaga gcagggtccc gcttccctaa
     2701 ggccctgctc tgggcttctg ggtttgagtc cttggcaagc ccaggagagg cgctcaggct
     2761 tccctgtccc ccttcctcgt ccaccatctc atgcccctgg ctctcctgcc ccttccctac
     2821 aggggttcct ggctctgctc tccaccgcgg tggagctcca gcttttgttc cctttagtga
     2881 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2941 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3001 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3061 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3121 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3181 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3241 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3301 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3361 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3421 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3481 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3541 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3601 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3661 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3721 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3781 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3841 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3901 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3961 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4021 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4081 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4141 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4201 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4261 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4321 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4381 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4441 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4501 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4561 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4621 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4681 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4741 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4801 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4861 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4921 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4981 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5041 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5101 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5161 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5221 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5281 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5341 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5401 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5461 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     5521 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5581 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     5641 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     5701 agggcgaatt gggtacctgc tttctctgac cagcattctc tcccctgggc ctgtgccgct
     5761 ttctgtctgc agcttgtggc ctgggtcacc tctacggctg gcccagatcc ttccctgccg
     5821 cctccttcag gttccgtctt cctccactcc ctcttcccct tgctctctgc tgtgttgctg
     5881 cccaaggatg ctctttccgg agcacttcct tctcggcgct gcaccacgtg atgtcctctg
     5941 agcggatcct ccccgtgtct gggtcctctc cgggcatctc tcctccctca cccaacccca
     6001 tgccgtcttc actcgctggg ttcccttttc cttctccttc tggggcctgt gccatctctc
     6061 gtttcttagg atggccttct ccgacggatg tctcccttgc gtcccgcctc cccttcttgt
     6121 aggcctgcat catcaccgtt tttctggaca accccaaagt accccgtctc cctggcttta
     6181 gccacctctc catcctcttg ctttctttgc ctggacaccc cgttctcctg tggattcggg
     6241 tcacctctca ctcctttcat ttgggcagct cccctacccc ccttacctct ctagtctgtg
     6301 ctagctcttc cagccccctg tcatggcatc ttccaggggt ccgagagctc agctagtctt
     6361 cttcctccaa cccgggcccc tatgtccact tcaggacagc atgtttgctg cctccaggga
     6421 tcctgtgtcc ccgagctggg accaccttat attcccaggg ccggttaatg tggctctggt
     6481 tctgggtact tttatctgtc ccctccaccc cacagtgggg caagcttatt accctgttat
     6541 ccctaaccgg tgcgttttta tgcttgtagt attgtataat gtttttaaga tccaagcttc
     6601 tgacctcttc tcttcctccc acagggcctc gagacaattg attaacatcg atacggtacc
     6661 gagttggcgc gcctgggagc tcacggggac agcccccccc caaagccccc agggatgtaa
     6721 ttacgtccct cccccgctag ggggcagcag cgagccgccc ggggctccgc tccggtccgg
     6781 cgctcccccc gcatccccga gccggcagcg tgcggggaca gcccgggcac ggggaaggtg
     6841 gcacgggatc gctttcctct gaacgcttct cgctgctctt tgagcctgca gacacctggg
     6901 gggatacggg gaaaaagctt taggctgaaa gagagattta gaatgacagg cgcgcctggc
     6961 catacatcga tacggtaccg agttggcgcg cctgggagct cacggggaca gccccccccc
     7021 aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg
     7081 gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag
     7141 cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt
     7201 gagcctgcag acacctgggg ggatacgggg aaaaagcttt aggctgaaag agagatttag
     7261 aatgacaggc gcgcctggtg gcgaacgat
//
