LOCUS       40924_8076        8833 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCAG-TagBFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8833)
  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  JOURNAL   Unpublished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 8833)
  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 8833)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8833)
  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            "Unpublished"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..8833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(222..810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(984..1841)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1842..1946)
                     /label=AmpR promoter
     rep_origin      complement(1972..2427)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2569..2585
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2595..2613
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2628..3431
                     /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    3510..3535
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    4207..4227
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     enhancer        4313..4616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4619..4894
                     /label=chicken beta-actin promoter
     intron          4900..5908
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     protein_bind    5966..5990
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6008..6697
                     /codon_start=1
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGH"
     misc_feature    6712..6733
                     /label=FF5
                     /note="FF5"
     misc_feature    6734..6755
                     /label=FF5
                     /note="FF5"
     misc_feature    6756..6777
                     /label=FF5
                     /note="FF5"
     misc_feature    6778..6799
                     /label=FF5
                     /note="FF5"
     protein_bind    complement(6866..6890)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    7045..7100
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7461..7477)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7485..7501
                     /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(7509..7539)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7554..7575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    7748..8584
                     /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(8614..8632)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8653..8669)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8677..8693)
                     /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(8701..8731)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(8746..8767)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
       61 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      121 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      181 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      241 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      301 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      361 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      421 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      481 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      541 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      601 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      661 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      721 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      781 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      841 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      901 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      961 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1021 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1081 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1141 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1201 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1261 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1321 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1381 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1441 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1501 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1561 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1621 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1681 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1741 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1801 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1861 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1921 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     1981 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2041 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2101 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2161 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2221 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2281 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2341 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2401 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     2461 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     2521 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     2581 ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtacctgc tttctctgac
     2641 cagcattctc tcccctgggc ctgtgccgct ttctgtctgc agcttgtggc ctgggtcacc
     2701 tctacggctg gcccagatcc ttccctgccg cctccttcag gttccgtctt cctccactcc
     2761 ctcttcccct tgctctctgc tgtgttgctg cccaaggatg ctctttccgg agcacttcct
     2821 tctcggcgct gcaccacgtg atgtcctctg agcggatcct ccccgtgtct gggtcctctc
     2881 cgggcatctc tcctccctca cccaacccca tgccgtcttc actcgctggg ttcccttttc
     2941 cttctccttc tggggcctgt gccatctctc gtttcttagg atggccttct ccgacggatg
     3001 tctcccttgc gtcccgcctc cccttcttgt aggcctgcat catcaccgtt tttctggaca
     3061 accccaaagt accccgtctc cctggcttta gccacctctc catcctcttg ctttctttgc
     3121 ctggacaccc cgttctcctg tggattcggg tcacctctca ctcctttcat ttgggcagct
     3181 cccctacccc ccttacctct ctagtctgtg ctagctcttc cagccccctg tcatggcatc
     3241 ttccaggggt ccgagagctc agctagtctt cttcctccaa cccgggcccc tatgtccact
     3301 tcaggacagc atgtttgctg cctccaggga tcctgtgtcc ccgagctggg accaccttat
     3361 attcccaggg ccggttaatg tggctctggt tctgggtact tttatctgtc ccctccaccc
     3421 cacagtgggg caagcttatt accctgttat ccctaaccgg tctaataccc aatctctcgt
     3481 cttatccaga tgttttatac gccaagcttc tgacctcttc tcttcctccc acagggcctc
     3541 gagacaattg catcgatggt accagtgaaa gtctgcaatg aattcgagtt ggcgcgcctg
     3601 tcattctaaa tctctctttc agcctaaagc tttttccccg tatcccccca ggtgtctgca
     3661 ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc
     3721 ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg accggagcgg
     3781 agccccgggc ggctcgctgc tgccccctag cgggggaggg acgtaattac atccctgggg
     3841 gctttgggag ggggctgtcc ccgtgagctc ccaggcgcgc ctgtcattct aaatctctct
     3901 ttcagcctaa agctttttcc ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga
     3961 gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct gtccccgcac
     4021 gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg ggcggctcgc
     4081 tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg gggggggctg
     4141 tccccgtgag ctcccaggcg cgccaactcg gtaccgtatc gatgtcgacg ttaacgctag
     4201 tgatatcaac tttgtataga aaagttggct ccgaatttct cgagttaatt aatagttatt
     4261 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     4321 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     4381 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     4441 actatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     4501 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4561 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatgggtc
     4621 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     4681 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg ggggggggcg
     4741 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
     4801 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
     4861 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gagtcgctgc gttgccttcg
     4921 ccccgtgccc cgctccgcgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
     4981 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
     5041 taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc
     5101 cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
     5161 cgcgtgcggc ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
     5221 gcgctccgcg tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     5281 ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt
     5341 gggcgcggcg gtcgggctgt aacccccccc tgcacccccc tccccgagtt gctgagcacg
     5401 gcccggcttc gggtgcgggg ctccgtgcgg ggcgtggcgc ggggctcgcc gtgccgggcg
     5461 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     5521 cgggggaggg gcgcggcggc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag
     5581 ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg
     5641 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg gcgaagcggt
     5701 gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc
     5761 cccttctcca tctccagcct cggggctgcc gcagggggac ggctgccttc gggggggacg
     5821 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
     5881 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggttg ttgtgctgtc
     5941 tcatcatttt ggcaaagaat tcgacccaag tttgtacaaa aaagcaggct gattaccggt
     6001 cgccaccatg agcgagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac
     6061 cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
     6121 ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct
     6181 ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg gcatccccga
     6241 cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga
     6301 cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa
     6361 cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga agaaaacact
     6421 cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg aaggcagaaa
     6481 cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacatca agaccacata
     6541 tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg tggactacag
     6601 actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg aggtggcagt
     6661 ggccagatac tgcgacctcc ctagcaaact ggggcactaa gcggccgcaa aaagcactct
     6721 gatttgacaa ttaaagcact ctgatttgac aattaaagca ctctgatttg acaattaaag
     6781 cactctgatt tgacaattaa agcttcgaat tctgcagtcg acggtaccgc gggcccggga
     6841 tccaccggat ctagataact gatctaccca gctttcttgt acaaagtggt acgcgtgaat
     6901 tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg
     6961 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     7021 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     7081 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     7141 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     7201 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     7261 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     7321 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     7381 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc
     7441 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     7501 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     7561 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     7621 tgccagcgga tcgacagtac taagcttggt gaattccctt atgtgagtgt aaaaggcagg
     7681 cgagtttgtc ccttaattaa tagggataac agggtaatct cgaggtttaa acgtcgacgc
     7741 ggccgctact agggacagga ttggtgacag aaaagcccca tccttaggcc tcctccttcc
     7801 tagtctcctg atattgggtc taacccccac ctcctgttag gcagattcct tatctggtga
     7861 cacaccccca tttcctggag ccatctctct ccttgccaga acctctaagg tttgcttacg
     7921 atggagccag agaggatcct gggagggaga gcttggcagg gggtgggagg gaaggggggg
     7981 atgcgtgacc tgcccggttc tcagtggcca ccctgcgcta ccctctccca gaacctgagc
     8041 tgctctgacg cggctgtctg gtgcgtttca ctgatcctgg tgctgcagct tccttacact
     8101 tcccaagagg agaagcagtt tggaaaaaca aaatcagaat aagttggtcc tgagttctaa
     8161 ctttggctct tcacctttct agtccccaat ttatattgtt cctccgtgcg tcagttttac
     8221 ctgtgagata aggccagtag ccagccccgt cctggcaggg ctgtggtgag gaggggggtg
     8281 tccgtgtgga aaactccctt tgtgagaatg gtgcgtccta ggtgttcacc aggtcgtggc
     8341 cgcctctact ccctttctct ttctccatcc ttctttcctt aaagagtccc cagtgctatc
     8401 tgggacatat tcctccgccc agagcagggt cccgcttccc taaggccctg ctctgggctt
     8461 ctgggtttga gtccttggca agcccaggag aggcgctcag gcttccctgt cccccttcct
     8521 cgtccaccat ctcatgcccc tggctctcct gccccttccc tacaggggtt cctggctctg
     8581 ctctccaccg cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     8641 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     8701 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     8761 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     8821 gcattaatga atc
//
