LOCUS       40924_11920        7657 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCMV-TALER2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7657)
  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  JOURNAL   Unpublished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 7657)
  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 7657)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7657)
  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            "Unpublished"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..7657
                     /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    1395..4220
                     /label=similar to TALER2
                     /note="similar to TALER2"
     CDS             4230..4250
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     polyA_signal    4445..4566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4573..5028)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5055..5159
                     /label=AmpR promoter
     promoter        5161..5518
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5553..6344
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    6579..6626
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6955..7543
                     /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 gaccattcgt ccgcgcaggc caagtcctgc ccgcgagctt ctgcccggac
     1441 cccaaccgga tagggttcag ccgactgcag atcgtggggt gtctgcgcct gctggcagcc
     1501 ctctggatgg cttgcccgct cggcggacgg tgtcccggac ccggctgcca tctccccctg
     1561 cgccctcacc tgcgttctcg gcgggcagct tcagcgatct gctccgtccg ttcgatccgt
     1621 cgcttcttga tacatcgctt cttgattcga tgcctgccgt cggcacgccg catacagcgg
     1681 ctgccccagc agagtgggat gaggcgcaat cggctctgcg tgcagccgat gacccgccac
     1741 ccaccgtgcg tgtcgctgtc actgccgcgc ggccgccgcg cgccaagccg gccccgcgac
     1801 ggcgtgctgc gcaaccctcc gacgcttcgc cggccgcgca ggtggatcta cgcacgctcg
     1861 gctacagtca gcagcagcaa gagaagatca aaccgaaggt gcgttcgaca gtggcgcagc
     1921 accacgaggc actggtgggc catgggttta cacacgcgca catcgttgcg ctcagccaac
     1981 acccggcagc gttagggacc gtcgctgtca cgtatcagca cataatcacg gcgttgccag
     2041 aggcgacaca cgaagacatc gttggcgtcg gcaaacagtg gtccggcgca cgcgccctgg
     2101 aggccttgct cacggatgcg ggggagttga gaggtccgcc gttacagttg gacacaggcc
     2161 aacttgtgaa gattgcaaaa cgtggcggcg tgaccgcaat ggaggcagtg catgcatcgc
     2221 gcaatgcact gacgggtgcc cccctgaacc tgaccccgga ccaagtggtg gctatcgcca
     2281 gcaacattgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc
     2341 aggaccatgg cctgaccccg gaccaagtgg tggctatcgc cagcaacatt ggcggcaagc
     2401 aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc
     2461 cggaccaagt ggtggctatc gccagcaaca atggcggcaa gcaagcgctc gaaacggtgc
     2521 agcggctgtt gccggtgctg tgccaggacc atggcctgac cccggaccaa gtggtggcta
     2581 tcgccagcaa cattggcggc aagcaagcgc tcgaaacggt gcagcggctg ttgccggtgc
     2641 tgtgccagga ccatggcctg accccggacc aagtggtggc tatcgccagc aacattggcg
     2701 gcaagcaagc gctcgaaacg gtgcagcggc tgttgccggt gctgtgccag gaccatggcc
     2761 tgaccccgga ccaagtggtg gctatcgcca gcaacggtgg cggcaagcaa gcgctcgaaa
     2821 cggtgcagcg gctgttgccg gtgctgtgcc aggaccatgg cctgaccccg gaccaagtgg
     2881 tggctatcgc cagcaacatt ggcggcaagc aagcgctcga aacggtgcag cggctgttgc
     2941 cggtgctgtg ccaggaccat ggcctgaccc cggaccaagt ggtggctatc gccagcaaca
     3001 ttggcggcaa gcaagcgctc gaaacggtgc agcggctgtt gccggtgctg tgccaggacc
     3061 atggcctgac cccggaccaa gtggtggcta tcgccagcaa cattggcggc aagcaagcgc
     3121 tcgaaacggt gcagcggctg ttgccggtgc tgtgccagga ccatggcctg accccggacc
     3181 aagtggtggc tatcgccagc aacggtggcg gcaagcaagc gctcgaaacg gtgcagcggc
     3241 tgttgccggt gctgtgccag gaccatggcc tgaccccgga ccaagtggtg gctatcgcca
     3301 gcaacattgg cggcaagcaa gcgctcgaaa cggtgcagcg gctgttgccg gtgctgtgcc
     3361 aggaccatgg cctgaccccg gaccaagtgg tggctatcgc cagcaacggt ggcggcaagc
     3421 aagcgctcga aacggtgcag cggctgttgc cggtgctgtg ccaggaccat ggcctgaccc
     3481 cggaccaagt ggtggctatc gccagcaaca ttggcggcaa gcaagcgctc gaaagcattg
     3541 tggcccagct gagccggcct gatccggcgt tggccgcgtt gaccaacgac cacctcgtcg
     3601 ccttggcctg cctcggcgga cgtcctgcca tggatgcagt gaaaaaggga ttgccgcacg
     3661 cgccggaatt gatcagaaga gtcaatcgcc gtattggcga acgcacgtcc catcgcgttg
     3721 ccgactacgc gcaagtggtt cgcgtgctgg agtttttcca gtgccactcc cacccagcgt
     3781 acgcatttga tgaggccatg acgcagttcg ggatgagcag gaacgggttg gtacagctct
     3841 ttcgcagagt gggcgtcacc gaactcgaag cccgcggtgg aacgctcccc ccagcctcgc
     3901 agcgttggga ccgtatcctc caggcatcag ggatgaaaag ggccaaaccg tcccctactt
     3961 cagctcaaac accggatcag gcgtctttgc atgcattcgc cgattcgctg gagcgtgacc
     4021 ttgatgcgcc cagcccaatg cacgagggag atcagacgcg ggcaagcagc cgtaaacggt
     4081 cccgatcgga tcgtgctgtc accggcccct ccgcacagca ggctgtcgag gtgcgcgttc
     4141 ccgaacagcg cgatgcgctg catttgcccc tcagctggag ggtaaaacgc ccgcgtacca
     4201 ggatctgggg cggcctcccg gatcccagcc ccaagaagaa gagaaaggtg gaggccagcg
     4261 gtggcggctc aaagcttggt ggcggctcaa ctagttaagg gcccggcgcg cctaaggtac
     4321 ccccgggtct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     4381 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     4441 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4501 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4561 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4621 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4681 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4741 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4801 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4861 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4921 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4981 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     5041 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5101 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     5161 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5221 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     5281 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5341 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5401 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5461 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     5521 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5581 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5641 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5701 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5761 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5821 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5881 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5941 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6001 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6061 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6121 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6181 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6241 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6301 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6361 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6421 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6481 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     6541 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6601 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6661 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6721 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6781 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6841 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6901 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6961 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7021 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7081 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     7141 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7201 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7261 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7321 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7381 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7441 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7501 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7561 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7621 ccctgattct gtggataacc gtattaccgc catgcat
//
