LOCUS       Exported                8300 bp ds-DNA     circular SYN 02-FEB-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8300)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2018 from SnapGene 4.0.8
            http://www.snapgene.com
COMMENT     LOCUS 055_PB_PGK_H2B_m 7686 bp ds-DNA circular 14-AUG-2013 
            DEFINITION . ACCESSION urn.local...1367881019035.10 (modified) 
            KEYWORDS . SOURCE ORGANISM . . COMMENT COMMENT ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     source          1..8300
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1944..2411
     source          2277..2372
     primer_bind     812..831
                     /label=BII(R)
     repeat_region   874..936
                     /label=5'TR
     repeat_region   887..917
                     /label=5'TR(1)
     misc_feature    887..917
                     /label=31bp spacer
     enhancer        1112..1343
                     /label=core insul
     promoter        1357..1362
                     /label=CMV7 promoter
     primer_bind     1402..1407
                     /label=5'TREtight MluI
     misc_feature    1408..1921
                     /label=PGK promoter
                     /note="Geneious type: Promoter Eukaryotic"
     primer_bind     1408..1427
                     /label=forward primer
     primer_bind     1930..1937
                     /label=primer3
                     /note="Geneious name: Primer"
     misc_feature    1930..1937
                     /label=Kozak
                     /note="Kozak"
     CDS             1944..2264
                     /codon_start=1
                     /label=FKBP12 F36V
                     /note="FKBP12 F36V"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KPE"
     CDS             2277..2366
                     /codon_start=1
                     /label=CZp
                     /translation="EQLEKKLQALEKKLAQLEWKNQALEKKLAQ"
     CDS             2382..2402
                     /codon_start=1
                     /label=TEVs
                     /note="TEVs"
                     /translation="ENLYFQS"
     CDS             2415..3128
                     /codon_start=1
                     /label=Citrine
                     /note="Citrine"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             3144..3164
                     /codon_start=1
                     /label=TVMVsite
                     /note="TVMVsite"
                     /translation="ETVRFQS"
     CDS             3177..3650
                     /codon_start=1
                     /label=DHFR
                     /note="DHFR"
                     /translation="ISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     misc_feature    3673..3674
                     /label=GC
     polyA_signal    3679..3893
                     /label=BGH polyadenylation sequence
     primer_bind     3798..3821
                     /label=1,857 F
     misc_feature    3893..3894
                     /label=GATC
                     /note="Geneious type: Editing History Deletion"
     misc_feature    3970..4291
                     /label=SV40 promoter and origin
                     /note="Geneious type: promoter uukaryotic"
     misc_feature    4307..4373
                     /label=EM-7 promoter
                     /note="Geneious type: promoter uukaryotic"
     CDS             4374..4748
                     /label=Zeocin resistance gene
     primer_bind     4477..4493
                     /label=2,540 F
     misc_feature    4755..4859
                     /label=WPRE
     misc_feature    4865..5000
                     /label=SV40 pA
     polyA_signal    4870..4999
                     /label=SV40 polyadenylation sequence
     enhancer        complement(5249..5479)
                     /label=core insul(1)
     repeat_region   5754..5788
                     /label=3'TR
     primer_bind     complement(6307..6326)
                     /label=pLac(F)
     CDS             complement(7440..8300)
                     /label=Ampicillin resistance gene
     CDS             complement(7443..8300)
                     /label=Amp
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccacta gtaggcgttt tgcgctgctt
     1381 cgcgatgtac gggccagata tacgcgttct accgggtagg ggaggcgctt ttcccaaggc
     1441 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     1501 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
     1561 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
     1621 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcat tagtctcgtg cagatggaca
     1681 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
     1741 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
     1801 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
     1861 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca
     1921 gcccaagctt cgccaccatg ggaggagttc aggtggagac catatctccg ggggatggca
     1981 ggacatttcc gaaaaggggc cagacgtgtg ttgttcatta tacagggatg ttggaggacg
     2041 gaaaaaaggt agactcttct cgggatagaa ataagccctt taagttcatg ctgggtaagc
     2101 aggaagttat tcgaggatgg gaagaaggag tagcccagat gtcagttgga cagcgggcaa
     2161 aattgactat aagccctgat tatgcctacg gtgcaacggg tcacccaggt attataccgc
     2221 cgcatgcgac actcgttttt gacgtggaac ttttgaaacc tgaaggaggc ggatccgagc
     2281 aactcgaaaa gaagctgcaa gcgttggaga agaagctcgc acaacttgaa tggaaaaatc
     2341 aggcactgga aaagaaactg gctcaaggtg gctccggcag cgagaatctt tatttccaat
     2401 ccggctcagg ctccgtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2461 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2521 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     2581 cctggcccac cctcgtgacc accttcggct acggcctgat gtgcttcgcc cgctaccccg
     2641 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2701 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2761 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2821 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     2881 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     2941 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     3001 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg
     3061 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     3121 tgtacaaggg tgggtccggt ctcgagacgg ttagattcca gagtgggagc ggatcgatat
     3181 cactgatcgc tgcactcgcg gttgattatg taataggcat ggaaaacgcg atgccctgga
     3241 acctgccagc agacctcgca tggtttaagc ggaacactct gaataagcca gtcattatgg
     3301 gccgacatac gtgggaaagc attggacgcc cactgccagg aaggaagaat ataatcctgt
     3361 catcacagcc gtccacggac gatagagtca cttgggtaaa atccgttgac gaagccatag
     3421 ctgcctgtgg tgatgtgcct gagataatgg taataggagg agggcgggta atcgaacaat
     3481 ttctgcctaa agcccagaag ctgtacctta cccatattga cgcggaagta gagggcgaca
     3541 cacatttccc agattatgag ccagatgatt gggagagcgt tttctccgaa tttcatgatg
     3601 cagacgccca gaatagccac agctactgct ttgaaatatt ggagcgccga tgaagctagc
     3661 gaattcgaat ttaaatcggc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3721 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3781 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3841 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggacgcggcc
     3901 gcaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4021 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4081 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4141 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4201 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4261 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     4321 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     4381 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     4441 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     4501 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     4561 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     4621 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     4681 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     4741 aggactgagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     4801 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     4861 cgttaactaa acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4921 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4981 aatgtatctt atcatgtctg gaattgactc aaatgatgtc aattagtcta tcagaagctc
     5041 atctggtctc ccttccgggg gacaagacat ccctgtttaa tatttaaaca gcagtgttcc
     5101 caaactgggt tcttatatcc cttgctctgg tcaaccaggt tgcagggttt cctgtcctca
     5161 caggaacgaa gtccctaaag aaacagtggc agccaggttt agccccggaa ttgactggat
     5221 tcctttttta gggcccattg gtatggcttt ttccccgtat ccccccaggt gtctgcaggc
     5281 tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg
     5341 ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc ggagcggagc
     5401 cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc cctgggggct
     5461 ttgggggggg gctgtccctg atatctataa caagaaaata tatatataat aagttatcac
     5521 gtaagtagaa catgaaataa caatataatt atcgtatgag ttaaatctta aaagtcacgt
     5581 aaaagataat catgcgtcat tttgactcac gcggtcgtta tagttcaaaa tcagtgacac
     5641 ttaccgcatt gacaagcacg cctcacggga gctccaagcg gcgactgaga tgtcctaaat
     5701 gcacagcgac ggattcgcgc tatttagaaa gagagagcaa tatttcaaga atgcatgcgt
     5761 caattttacg cagactatct ttctagggtt aatctagctg catcaggatc atatcgtcgg
     5821 gtcttttttc cggctcagtc atcgcccaag ctggcgctat ctgggcatcg gggaggaaga
     5881 agcccgtgcc ttttcccgcg aggttgaagc ggcatggaaa gagtttgccg aggatgactg
     5941 ctgctgcatt gacgttgagc gaaaacgcac gtttaccatg atgattcggg aaggtgtggc
     6001 catgcacgcc tttaacggtg aactgttcgt tcaggccacc tgggatacca gttcgtcgcg
     6061 gcttttccgg acacagttcc ggatggtcag cccgaagcgc atcagcaacc cgaacaatac
     6121 cggcgacagc cggaactgcc gtgccggtgt gcagattaat gacagcggtg cggcgctggg
     6181 atattacgtc agcgaggacg ggtatcctgg ctggatgccg cagaaatgga catggatacc
     6241 ccgtgagtta cccggcgggc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     6301 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6361 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6421 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6481 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6541 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6601 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6661 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6721 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6781 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6841 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6901 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6961 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7021 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7081 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     7141 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7201 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7261 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7321 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7381 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7441 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7501 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7561 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7621 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7681 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7741 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7801 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7861 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7921 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7981 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8041 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8101 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8161 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8221 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8281 cggaaatgtt gaatactcat
//
