LOCUS       40924_11935        7423 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCMV-TALER22, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7423)
  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  JOURNAL   Unpublished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 7423)
  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 7423)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7423)
  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            "Unpublished"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..7423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             603..1319
                     /codon_start=1
                     /label=mEYFP
                     /note="enhanced YFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1326..1379
                     /label=2A
                     /note="2A"
     CDS             1326..1379
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    1394..3985
                     /label=similar to TALER22
                     /note="similar to TALER22"
     CDS             3996..4016
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     polyA_signal    4211..4332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4339..4794)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4821..4925
                     /label=AmpR promoter
     promoter        4927..5284
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5319..6110
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    6345..6392
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6721..7309
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgaat
      601 tcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      661 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      721 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      781 ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
      841 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      901 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      961 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1021 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1081 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1141 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1201 actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     1261 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc
     1321 aattggaggg ccgcggcagc ctgctgacct gcggcgacgt ggaggaaaac cccggcccca
     1381 ccggtttgtc gacgtggacc tgagaacact gggatattct cagcagcagc aggagaagat
     1441 caagcccaag gtgagatcca cagtggccca gcaccacgaa gccctggtgg gacacggatt
     1501 tacacacgcc cacattgtgg ccctgtctca gcaccctgcc gccctgggaa cagtggccgt
     1561 gaaatatcag gatatgattg ccgccctgcc tgaggccaca cacgaagcca ttgtgggagt
     1621 gggaaaacag tggtctggag ccagagccct ggaagccctg ctgacagtgg ccggagaact
     1681 gagaggacct cctctgcagc tggatacagg acagctgctg aagattgcca aaaggggcgg
     1741 agtgaccgcg gtggaagccg tgcacgcctg gagaaatgcc ctgacaggag cccctctgaa
     1801 cctgaccccc gaacaggtgg tggccattgc cagcaacatc ggcggcaagc aggccctgga
     1861 aaccgtgcag agactgctgc ccgtgctgtg ccaggcccat ggcctgacac ctgaacaggt
     1921 ggtggctatc gcctctaaca agggaggaaa acaggctctg gaaacagtgc agcggctgct
     1981 gcctgtgctg tgtcaggctc acggcttgac tccagaacag gtggtggcta ttgcttccaa
     2041 cggcgggggg aaacaggccc tggaaactgt gcagcgcctg ctgccagtgc tgtgccaggc
     2101 tcacggactg acccccgaac aggtggtggc cattgccagc cacgacggcg gcaagcaggc
     2161 cctggaaacc gtgcagagac tgctgcccgt gctgtgccag gcccatggcc tgacacctga
     2221 acaggtggtg gctatcgcct ctaatatcgg aggaaaacag gctctggaaa cagtgcagcg
     2281 gctgctgcct gtgctgtgtc aggctcacgg cttgactcca gaacaggtgg tggctattgc
     2341 ttccaacaag ggggggaaac aggccctgga aactgtgcag cgcctgctgc cagtgctgtg
     2401 ccaggctcac gggctgaccc ccgaacaggt ggtggccatt gccagccacg acggcggcaa
     2461 gcaggccctg gaaaccgtgc agagactgct gcccgtgctg tgccaggccc atggcctgac
     2521 acctgaacag gtggtggcta tcgcctctaa tatcggagga aaacaggctc tggaaacagt
     2581 gcagcggctg ctgcctgtgc tgtgtcaggc tcacggcttg actccagaac aggtggtggc
     2641 tattgcttcc cacgacgggg ggaaacaggc cctggaaact gtgcagcgcc tgctgccagt
     2701 gctgtgccag gctcacggcc tcactcccga acaggtggtg gccattgcca gccacgacgg
     2761 cggcaagcag gccctggaaa ccgtgcagag actgctgccc gtgctgtgcc aggcccatgg
     2821 cctgacacct gaacaggtgg tggctatcgc ctctaatatc ggaggaaaac aggctctgga
     2881 aacagtgcag cggctgctgc ctgtgctgtg tcaggctcac ggcttgactc cagaacaggt
     2941 ggtggctatt gcttccaaca agggggggaa acaggccctg gaaactgtgc agcgcctgct
     3001 gccagtgctg tgccaggctc acggactgac ccccgaacag gtggtggcca ttgccagcaa
     3061 caagggcggc aagcaggccc tggaaaccgt gcagagactg ctgcccgtgc tgtgccaggc
     3121 ccatggcctg acacctgaac aggtggtggc tatcgcctct cacgacggag gaaaacaggc
     3181 tctggaaaca gtgcagcggc tgctgcctgt gctgtgtcag gctcacggct tgactccaga
     3241 acaggtggtg gctattgctt ccaatattgg ggggaaacag gccctggaaa ctgtgcagcg
     3301 cctgctgcca gtgctgtgcc aggctcacgg gctgaccccc gaacaggtgg tggccattgc
     3361 cagcaacggc ggcggcaagc aggccctgga aaccgtgcag agactgctgc ccgtgctgtg
     3421 ccaggcccat ggcctgacac ctgaacaggt ggtggctatc gcctctaacg gcggaggaaa
     3481 acaggctctg gaaacagtgc agcggctgct gcctgtgctg tgtcaggctc acggcttgac
     3541 tccacagcag gtcgtggcaa ttgctagcaa cggcggcgga cggcccgccc tggagagcat
     3601 tgtggcccag ctgtctagac ctgatcctgc cctggccgcc ctgacaaatg atcacctggt
     3661 ggccctggcc tgtctgggag gcagacctgc cctggatgcc gtgaaaaaag gactgcctca
     3721 cgcccctgcc ctgatcaaga gaacaaatag aagaatcccc gagcggacct ctcacagagt
     3781 ggccgatcac gcccaggtgg tgagagtgct gggatttttt cagtgtcact ctcaccctgc
     3841 ccaggccttt gatgatgcca tgacacagtt tggcatgagc agacacggac tgctgcagct
     3901 gtttagaaga gtgggagtga cagaactgga ggccagatcc ggaaccctgc ctcctgcctc
     3961 tcagagatgg gataggattc tgcagggatc ccagccccaa gaagaagaga aaggtggagg
     4021 ccagcggtgg cggctcaaag cttggtggcg gctcaactag ttaagggccc ggcgcgccta
     4081 aggtaccccc gggtctagat aactgatcat aatcagccat accacatttg tagaggtttt
     4141 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4201 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4261 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4321 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4381 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4441 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4501 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4561 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4621 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4681 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4741 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4801 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4861 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4921 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4981 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5041 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5101 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5161 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5221 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5281 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5341 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5401 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5461 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     5521 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5581 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5641 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5701 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5761 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5821 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5881 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5941 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6001 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6061 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6121 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6181 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6241 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     6301 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     6361 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     6421 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     6481 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     6541 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6601 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6661 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6721 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6781 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6841 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     6901 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6961 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7021 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7081 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7141 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7201 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7261 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7321 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7381 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
