LOCUS       40924_11945        9605 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCMV-TALER24, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9605)
  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  JOURNAL   Unpublished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 9605)
  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 9605)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9605)
  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            "Unpublished"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..9605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..21
                     /label=attB4
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     enhancer        93..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    631..655
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      664..673
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             673..714
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     CDS             727..747
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     misc_feature    757..3348
                     /label=similar to TALER24
                     /note="similar to TALER24"
     protein_bind    complement(3439..3463)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3618..3673
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4034..4050)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4058..4074
                     /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(4082..4112)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4127..4148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    4321..5157
                     /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(5187..5205)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5226..5242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5250..5266)
                     /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(5274..5304)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5319..5340)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5628..6216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6390..7247)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7248..7352)
                     /label=AmpR promoter
     rep_origin      complement(7378..7833)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     7975..7991
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8001..8019
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    8034..8837
                     /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    8916..8941
                     /label=SA
                     /note="splice acceptor site"
ORIGIN
        1 caactttgta tagaaaagtt ggctccgaat tctagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 ggtttagtga accgtcagat cgaattcgac ccaagtttgt acaaaaaagc aggctgcggc
      661 cgcgccacca tgggaaaacc tattcctaat cctctgctgg gcctggattc taccggaggc
      721 atggccccta agaaaaagcg gaaggtggac ggcggagtgg acctgagaac actgggatat
      781 tctcagcagc agcaggagaa gatcaagccc aaggtgagat ccacagtggc ccagcaccac
      841 gaagccctgg tgggacacgg atttacacac gcccacattg tggccctgtc tcagcaccct
      901 gccgccctgg gaacagtggc cgtgaaatat caggatatga ttgccgccct gcctgaggcc
      961 acacacgaag ccattgtggg agtgggaaaa cagtggtctg gagccagagc cctggaagcc
     1021 ctgctgacag tggccggaga actgagagga cctcctctgc agctggatac aggacagctg
     1081 ctgaagattg ccaaaagggg cggagtgacc gcggtggaag ccgtgcacgc ctggagaaat
     1141 gccctgacag gagcccctct gaacctgacc cccgaacagg tggtggccat tgccagcaac
     1201 atcggcggca agcaggccct ggaaaccgtg cagagactgc tgcccgtgct gtgccaggcc
     1261 catggcctga cacctgaaca ggtggtggct atcgcctcta acaagggagg aaaacaggct
     1321 ctggaaacag tgcagcggct gctgcctgtg ctgtgtcagg ctcacggctt gactccagaa
     1381 caggtggtgg ctattgcttc caatattggg gggaaacagg ccctggaaac tgtgcagcgc
     1441 ctgctgccag tgctgtgcca ggctcacgga ctgacccccg aacaggtggt ggccattgcc
     1501 agccacgacg gcggcaagca ggccctggaa accgtgcaga gactgctgcc cgtgctgtgc
     1561 caggcccatg gcctgacacc tgaacaggtg gtggctatcg cctctaatat cggaggaaaa
     1621 caggctctgg aaacagtgca gcggctgctg cctgtgctgt gtcaggctca cggcttgact
     1681 ccagaacagg tggtggctat tgcttccaat attgggggga aacaggccct ggaaactgtg
     1741 cagcgcctgc tgccagtgct gtgccaggct cacgggctga cccccgaaca ggtggtggcc
     1801 attgccagca acatcggcgg caagcaggcc ctggaaaccg tgcagagact gctgcccgtg
     1861 ctgtgccagg cccatggcct gacacctgaa caggtggtgg ctatcgcctc tcacgacgga
     1921 ggaaaacagg ctctggaaac agtgcagcgg ctgctgcctg tgctgtgtca ggctcacggc
     1981 ttgactccag aacaggtggt ggctattgct tcccacgacg gggggaaaca ggccctggaa
     2041 actgtgcagc gcctgctgcc agtgctgtgc caggctcacg gcctcactcc cgaacaggtg
     2101 gtggccattg ccagcaacat cggcggcaag caggccctgg aaaccgtgca gagactgctg
     2161 cccgtgctgt gccaggccca tggcctgaca cctgaacagg tggtggctat cgcctctcac
     2221 gacggaggaa aacaggctct ggaaacagtg cagcggctgc tgcctgtgct gtgtcaggct
     2281 cacggcttga ctccagaaca ggtggtggct attgcttcca atattggggg gaaacaggcc
     2341 ctggaaactg tgcagcgcct gctgccagtg ctgtgccagg ctcacggact gacccccgaa
     2401 caggtggtgg ccattgccag caacatcggc ggcaagcagg ccctggaaac cgtgcagaga
     2461 ctgctgcccg tgctgtgcca ggcccatggc ctgacacctg aacaggtggt ggctatcgcc
     2521 tctcacgacg gaggaaaaca ggctctggaa acagtgcagc ggctgctgcc tgtgctgtgt
     2581 caggctcacg gcttgactcc agaacaggtg gtggctattg cttccaacgg cggggggaaa
     2641 caggccctgg aaactgtgca gcgcctgctg ccagtgctgt gccaggctca cgggctgacc
     2701 cccgaacagg tggtggccat tgccagccac gacggcggca agcaggccct ggaaaccgtg
     2761 cagagactgc tgcccgtgct gtgccaggcc catggcctga cacctgaaca ggtggtggct
     2821 atcgcctcta acggcggagg aaaacaggct ctggaaacag tgcagcggct gctgcctgtg
     2881 ctgtgtcagg ctcacggctt gactccacag caggtcgtgg caattgctag caacggcggc
     2941 ggacggcccg ccctggagag cattgtggcc cagctgtcta gacctgatcc tgccctggcc
     3001 gccctgacaa atgatcacct ggtggccctg gcctgtctgg gaggcagacc tgccctggat
     3061 gccgtgaaaa aaggactgcc tcacgcccct gccctgatca agagaacaaa tagaagaatc
     3121 cccgagcgga cctctcacag agtggccgat cacgcccagg tggtgagagt gctgggattt
     3181 tttcagtgtc actctcaccc tgcccaggcc tttgatgatg ccatgacaca gtttggcatg
     3241 agcagacacg gactgctgca gctgtttaga agagtgggag tgacagaact ggaggccaga
     3301 tccggaaccc tgcctcctgc ctctcagaga tgggatagga ttctgcaggg ttcccgttta
     3361 aacaagcttg tcgacggtac cgaattcatc gatagtactc tcgagggatc cgagctcaag
     3421 atcttagcta agtagaccac ccagctttct tgtacaaagt ggtacgcgtg aattcactcc
     3481 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     3541 ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
     3601 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     3661 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
     3721 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
     3781 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
     3841 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
     3901 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
     3961 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
     4021 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4081 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4141 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4201 ggatcgacag tactaagctt ggtctaatac ccaatctctc gtcttatcca gatgttttat
     4261 acgccttaat taatagggat aacagggtaa tctcgaggtt taaacgtcga cgcggccgct
     4321 actagggaca ggattggtga cagaaaagcc ccatccttag gcctcctcct tcctagtctc
     4381 ctgatattgg gtctaacccc cacctcctgt taggcagatt ccttatctgg tgacacaccc
     4441 ccatttcctg gagccatctc tctccttgcc agaacctcta aggtttgctt acgatggagc
     4501 cagagaggat cctgggaggg agagcttggc agggggtggg agggaagggg gggatgcgtg
     4561 acctgcccgg ttctcagtgg ccaccctgcg ctaccctctc ccagaacctg agctgctctg
     4621 acgcggctgt ctggtgcgtt tcactgatcc tggtgctgca gcttccttac acttcccaag
     4681 aggagaagca gtttggaaaa acaaaatcag aataagttgg tcctgagttc taactttggc
     4741 tcttcacctt tctagtcccc aatttatatt gttcctccgt gcgtcagttt tacctgtgag
     4801 ataaggccag tagccagccc cgtcctggca gggctgtggt gaggaggggg gtgtccgtgt
     4861 ggaaaactcc ctttgtgaga atggtgcgtc ctaggtgttc accaggtcgt ggccgcctct
     4921 actccctttc tctttctcca tccttctttc cttaaagagt ccccagtgct atctgggaca
     4981 tattcctccg cccagagcag ggtcccgctt ccctaaggcc ctgctctggg cttctgggtt
     5041 tgagtccttg gcaagcccag gagaggcgct caggcttccc tgtccccctt cctcgtccac
     5101 catctcatgc ccctggctct cctgcccctt ccctacaggg gttcctggct ctgctctcca
     5161 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     5221 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5281 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5341 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5401 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5461 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5521 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5581 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5641 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5701 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5761 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5821 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5881 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5941 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6001 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6061 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6121 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6181 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6241 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6301 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6361 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6421 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6481 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6541 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6601 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6661 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6721 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6781 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6841 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6901 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6961 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7021 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7081 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7141 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7201 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7261 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7321 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     7381 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     7441 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     7501 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     7561 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     7621 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     7681 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     7741 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     7801 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     7861 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     7921 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     7981 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt acctgctttc
     8041 tctgaccagc attctctccc ctgggcctgt gccgctttct gtctgcagct tgtggcctgg
     8101 gtcacctcta cggctggccc agatccttcc ctgccgcctc cttcaggttc cgtcttcctc
     8161 cactccctct tccccttgct ctctgctgtg ttgctgccca aggatgctct ttccggagca
     8221 cttccttctc ggcgctgcac cacgtgatgt cctctgagcg gatcctcccc gtgtctgggt
     8281 cctctccggg catctctcct ccctcaccca accccatgcc gtcttcactc gctgggttcc
     8341 cttttccttc tccttctggg gcctgtgcca tctctcgttt cttaggatgg ccttctccga
     8401 cggatgtctc ccttgcgtcc cgcctcccct tcttgtaggc ctgcatcatc accgtttttc
     8461 tggacaaccc caaagtaccc cgtctccctg gctttagcca cctctccatc ctcttgcttt
     8521 ctttgcctgg acaccccgtt ctcctgtgga ttcgggtcac ctctcactcc tttcatttgg
     8581 gcagctcccc tacccccctt acctctctag tctgtgctag ctcttccagc cccctgtcat
     8641 ggcatcttcc aggggtccga gagctcagct agtcttcttc ctccaacccg ggcccctatg
     8701 tccacttcag gacagcatgt ttgctgcctc cagggatcct gtgtccccga gctgggacca
     8761 ccttatattc ccagggccgg ttaatgtggc tctggttctg ggtactttta tctgtcccct
     8821 ccaccccaca gtggggcaag cttattaccc tgttatccct aaccggtgcg tttttatgct
     8881 tgtagtattg tataatgttt ttaagatcca agcttctgac ctcttctctt cctcccacag
     8941 ggcctcgaga caattgatta acatcgatac ggtaccgagt tggcgcgcct gggagctcac
     9001 ggggacagcc cccccccaaa gcccccaggg atgtaattac gtccctcccc cgctaggggg
     9061 cagcagcgag ccgcccgggg ctccgctccg gtccggcgct ccccccgcat ccccgagccg
     9121 gcagcgtgcg gggacagccc gggcacgggg aaggtggcac gggatcgctt tcctctgaac
     9181 gcttctcgct gctctttgag cctgcagaca cctgggggga tacggggaaa aagctttagg
     9241 ctgaaagaga gatttagaat gacaggcgcg cctggccata catcgatacg gtaccgagtt
     9301 ggcgcgcctg ggagctcacg gggacagccc ccccccaaag cccccaggga tgtaattacg
     9361 tccctccccc gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc
     9421 cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg
     9481 ggatcgcttt cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat
     9541 acggggaaaa agctttaggc tgaaagagag atttagaatg acaggcgcgc ctggtggcga
     9601 acgat
//
