LOCUS       40924_11970        7380 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCMV-TALER32, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7380)
  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  JOURNAL   Unpublished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 7380)
  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 7380)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7380)
  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            "Unpublished"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..7380
                     /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    1386..3923
                     /label=similar to TALER32
                     /note="similar to TALER32"
     CDS             3930..3950
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     polyA_signal    4168..4289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4296..4751)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4778..4882
                     /label=AmpR promoter
     promoter        4884..5241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5276..6067
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    6302..6349
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6678..7266
                     /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 ccggtatgtc gcggacccgg ctcccttccc cacccgcacc cagcccagcg ttttcggccg
     1441 actcgttctc agacctgctt aggcagttcg acccctcact gtttaacaca tcgttgttcg
     1501 actcccttcc tccgtttggg gcgcaccata cggaggcggc caccggggag tgggatgagg
     1561 tgcagtcggg attgagagct gcggatgcac cacccccaac catgcgggtg gccgtcaccg
     1621 ctgcccgacc gccgagggcg aagcccgcac caaggcggag ggcagcgcaa ccgtccgacg
     1681 caagccccgc agcgcaagta gatttgagaa ctttgggata ttcacagcag cagcaggaaa
     1741 agatcaagcc caaagtgagg tcgacagtcg cgcagcatca cgaagcgctg gtgggtcatg
     1801 ggtttacaca tgcccacatc gtagccttgt cgcagcaccc tgcagccctt ggcacggtcg
     1861 ccgtcaagta ccaggacatg attgcggcgt tgccggaagc cacacatgag gcgatcgtcg
     1921 gtgtggggaa acagtggagc ggagcccgag cgcttgaggc cctgttgacg gtcgcgggag
     1981 agctgagagg gcctcccctt cagctggaca cgggccagtt gctgaagatc gcgaagcggg
     2041 gaggagtcac ggcggtcgag gcggtgcacg cgtggcgcaa tgcgctcacg ggagcacccc
     2101 tcaacctgac cccagagcag gtcgtggcaa tcgcctccaa cattggcggg aaacaggcac
     2161 tcgagactgt ccagcgcctg cttcccgtgc tgtgccaagc gcacggactc accccagagc
     2221 aggtcgtggc gatcgcaagc cacgacggag gaaagcaagc cttggaaaca gtacagaggc
     2281 tgttgcctgt gctgtgccaa gcgcacggcc tcaccccaga gcaggtcgtg gcaatcgcct
     2341 ccaacattgg cgggaaacag gcactcgaga ctgtccagcg cctgcttccc gtgctgtgcc
     2401 aagcgcacgg attaacccca gagcaggtcg tggcaatcgc ctccaacatt ggcgggaaac
     2461 aggcactcga gactgtccag cgcctgcttc ccgtgctgtg ccaagcgcac ggcttaaccc
     2521 cagagcaggt cgtggcgatc gcaagccacg acggaggaaa gcaagccttg gaaacagtac
     2581 agaggctgtt gcctgtgctg tgccaagcgc acggacttac cccagagcag gtcgtggcaa
     2641 tcgcctccaa cattggcggg aaacaggcac tcgagactgt ccagcgcctg cttcccgtgc
     2701 tgtgccaagc gcacggcctt accccagagc aggtcgtggc gatcgcaagc cacgacggag
     2761 gaaagcaagc cttggaaaca gtacagaggc tgttgcctgt gctgtgccaa gcgcacggac
     2821 taaccccaga gcaggtcgtg gcaatcgcct ccaacattgg cgggaaacag gcactcgaga
     2881 ctgtccagcg cctgcttccc gtgctgtgcc aagcgcacgg gctcacccca gagcaggtcg
     2941 tggcgatcgc aagccacgac ggaggaaagc aagccttgga aacagtacag aggctgttgc
     3001 ctgtgctgtg ccaagcgcac gggctaaccc cagagcaggt cgtggccatt gcctcgaatg
     3061 gagggggcaa acaggcgttg gaaaccgtac aacgattgct gccggtgctg tgccaagcgc
     3121 acggcctaac cccagagcag gtcgtggcaa tcgcctccaa cattggcggg aaacaggcac
     3181 tcgagactgt ccagcgcctg cttcccgtgc tgtgccaagc gcacgggtta accccagagc
     3241 aggtcgtggc cattgcctcg aatggagggg gcaaacaggc gttggaaacc gtacaacgat
     3301 tgctgccggt gctgtgccaa gcgcacggac tcacgcctga gcaggtagtg gctattgcat
     3361 ccaatatcgg gggcagaccc gcactggagt caatcgtggc ccagctttcg aggccggacc
     3421 ccgcgctggc cgcactcact aatgatcatc ttgtagcgct ggcctgcctc ggcggacgac
     3481 ccgccttgga tgcggtgaag aaggggctcc cgcacgcgcc tgcattgatt aagcggacca
     3541 acagaaggat tcccgagagg acatcacatc gagtggcaga tcacgcgcaa gtggtccgcg
     3601 tgctcggatt cttccagtgt cactcccacc ccgcacaagc gttcgatgac gccatgactc
     3661 aatttggtat gtcgagacac ggactgctgc agctctttcg tagagtcggt gtcacagaac
     3721 tcgaggcccg ctcgggcaca ctgcctcccg cctcccagcg gtgggacagg attctccaag
     3781 cgagcggtat gaaacgcgcg aagccttcac ctacgtcaac tcagacacct gaccaggcga
     3841 gccttcatgc gttcgcagac tcgctggaga gggatttgga cgcgccctcg cccatgcatg
     3901 aaggggacca aactcgcgcg tcagctagcc ccaagaagaa gagaaaggtg gaggccagcc
     3961 catggggcgg aggttcgaag cttggtggcg gctcaactag ttaagggccc ggcgcgccta
     4021 aggtaccccc gggtctagag atccaccgga tctagataac tgatcataat cagccatacc
     4081 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4141 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4201 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4261 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     4321 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4381 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4441 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4501 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4561 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4621 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4681 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4741 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4801 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4861 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     4921 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4981 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5041 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5101 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5161 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5221 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     5281 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5341 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5401 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     5461 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5521 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5581 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5641 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5701 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5761 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     5821 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5881 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5941 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     6001 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     6061 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6121 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6181 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6241 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     6301 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     6361 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     6421 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     6481 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     6541 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6601 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6661 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6721 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6781 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     6841 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6901 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6961 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7021 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7081 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7141 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7201 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7261 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7321 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
