LOCUS       40924_8026        6371 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCAG-EYFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6371)
  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  JOURNAL   Unpublished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 6371)
  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 6371)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6371)
  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            "Unpublished"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..6371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    112..233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(240..695)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        722..826
                     /label=AmpR promoter
     promoter        828..1185
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             1220..2011
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    2246..2293
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2622..3210
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     enhancer        3380..3683
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3686..3961
                     /label=chicken beta-actin promoter
     intron          3967..4975
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     CDS             5064..5780
                     /codon_start=1
                     /label=mEYFP
                     /note="enhanced YFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    5960..6081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
       61 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
      121 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
      181 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
      241 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
      301 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
      361 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
      421 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
      481 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
      541 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
      601 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
      661 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
      721 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
      781 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
      841 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
      901 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
      961 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     1021 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     1081 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     1141 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     1201 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     1261 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     1321 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     1381 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     1441 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     1501 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     1561 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     1621 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     1681 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     1741 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     1801 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     1861 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     1921 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     1981 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     2041 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     2101 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     2161 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     2221 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     2281 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     2341 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     2401 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     2461 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2521 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2581 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2641 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2701 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2761 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2821 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2881 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2941 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3001 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3061 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3121 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3181 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3241 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3301 gataaccgta tttaattaat agttattaat agtaatcaat tacggggtca ttagttcata
     3361 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3421 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     3481 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
     3541 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     3601 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     3661 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
     3721 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
     3781 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
     3841 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
     3901 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
     3961 gggagtcgag tcgctgcgtt gccttcgccc cgtgccccgc tccgcgccgc ctcgcgccgc
     4021 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
     4081 cctccgggct gtaattagcg cttggtttaa tgacggctcg tttcttttct gtggctgcgt
     4141 gaaagcctta aagggctccg ggagggccct ttgtgcgggg gggagcggct cggggggtgc
     4201 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggcccg cgctgcccgg cggctgtgag
     4261 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcgtgt gcgcgagggg agcgcggccg
     4321 ggggcggtgc cccgcggtgc gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     4381 gtgcgtgggg gggtgagcag ggggtgtggg cgcggcggtc gggctgtaac ccccccctgc
     4441 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtgcggggc
     4501 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     4561 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc ggagcgccgg
     4621 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     4681 gcagggactt cctttgtccc aaatctggcg gagccgaaat ctgggaggcg ccgccgcacc
     4741 ccctctagcg ggcgcgggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     4801 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccatct ccagcctcgg ggctgccgca
     4861 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     4921 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     4981 gggcaacgtg ctggttgttg tgctgtctca tcattttggc aaagaattcg gccattacgg
     5041 ccgctagctt accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     5101 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     5161 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     5221 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg
     5281 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     5341 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     5401 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     5461 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     5521 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     5581 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     5641 ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc aaagacccca
     5701 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     5761 gcatggacga gctgtacaag taagcggccg caaagcttcg aattctgcag tcgacggtac
     5821 cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     5881 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     5941 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     6001 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6061 caaactcatc aatgtatctt aacgcgttcg accgaataaa acctgtgacg gaagatcact
     6121 tcgcagaata aataaatcct ggtgtccctg ttgataccgg agatctccca cgccctgaag
     6181 ctgaaggacg gcggccacta cacctgcgac ttcaagaccg tgtacaaggc caagaagccc
     6241 gtgcagctgc ccggcaacca ctacgtggac tccaagctgg acatcaccaa ccacaacgag
     6301 gactacaccg tggtggagca gtacgagcac gccgaggccc gccactccgg ctcccagtag
     6361 agcggccgcg a
//
