LOCUS       V021062                 8929 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021062
VERSION     V021062
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             649..1365
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1399..3570
                     /label="PIK3R1(NM_181523)"
                     /note="PIK3R1(NM_181523)"
                     /gene="PIK3R1"
     CDS             3613..3678
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3705..4293
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4359..4407
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4609..5037
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5051..5380
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5447..6241
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6417..6538
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6587..6603)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6611..6627
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6635..6665)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6680..6701
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6989..7577)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7748..8608)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8609..8713)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgccaccat ggtgagcaag
      661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      781 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      841 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1081 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1141 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1201 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1261 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1321 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg agggggaagc
     1381 ggtaccgagc tcggatccat gagtgctgag gggtaccagt acagagcgct gtatgattat
     1441 aaaaaggaaa gagaagaaga tattgacttg cacttgggtg acatattgac tgtgaataaa
     1501 gggtccttag tagctcttgg attcagtgat ggacaggaag ccaggcctga agaaattggc
     1561 tggttaaatg gctataatga aaccacaggg gaaagggggg actttccggg aacttacgta
     1621 gaatatattg gaaggaaaaa aatctcgcct cccacaccaa agccccggcc acctcggcct
     1681 cttcctgttg caccaggttc ttcgaaaact gaagcagatg ttgaacaaca agctttgact
     1741 ctcccggatc ttgcagagca gtttgcccct cctgacattg ccccgcctct tcttatcaag
     1801 ctcgtggaag ccattgaaaa gaaaggtctg gaatgttcaa ctctatacag aacacagagc
     1861 tccagcaacc tggcagaatt acgacagctt cttgattgtg atacaccctc cgtggacttg
     1921 gaaatgatcg atgtgcacgt tttggctgac gctttcaaac gctatctcct ggacttacca
     1981 aatcctgtca ttccagcagc cgtttacagt gaaatgattt ctttagctcc agaagtacaa
     2041 agctccgaag aatatattca gctattgaag aagcttatta ggtcgcctag catacctcat
     2101 cagtattggc ttacgcttca gtatttgtta aaacatttct tcaagctctc tcaaacctcc
     2161 agcaaaaatc tgttgaatgc aagagtactc tctgaaattt tcagccctat gcttttcaga
     2221 ttctcagcag ccagctctga taatactgaa aacctcataa aagttataga aattttaatc
     2281 tcaactgaat ggaatgaacg acagcctgca ccagcactgc ctcctaaacc accaaaacct
     2341 actactgtag ccaacaacgg tatgaataac aatatgtcct tacaagatgc tgaatggtac
     2401 tggggagata tctcgaggga agaagtgaat gaaaaacttc gagatacagc agacgggacc
     2461 tttttggtac gagatgcgtc tactaaaatg catggtgatt atactcttac actaaggaaa
     2521 gggggaaata acaaattaat caaaatattt catcgagatg ggaaatatgg cttctctgac
     2581 ccattaacct tcagttctgt ggttgaatta ataaaccact accggaatga atctctagct
     2641 cagtataatc ccaaattgga tgtgaaatta ctttatccag tatccaaata ccaacaggat
     2701 caagttgtca aagaagataa tattgaagct gtagggaaaa aattacatga atataacact
     2761 cagtttcaag aaaaaagtcg agaatatgat agattatatg aagaatatac ccgcacatcc
     2821 caggaaatcc aaatgaaaag gacagctatt gaagcattta atgaaaccat aaaaatattt
     2881 gaagaacagt gccagaccca agagcggtac agcaaagaat acatagaaaa gtttaaacgt
     2941 gaaggcaatg agaaagaaat acaaaggatt atgcataatt atgataagtt gaagtctcga
     3001 atcagtgaaa ttattgacag tagaagaaga ttggaagaag acttgaagaa gcaggcagct
     3061 gagtatcgag aaattgacaa acgtatgaac agcattaaac cagaccttat ccagctgaga
     3121 aagacgagag accaatactt gatgtggttg actcaaaaag gtgttcggca aaagaagttg
     3181 aacgagtggt tgggcaatga aaacactgaa gaccaatatt cactggtgga agatgatgaa
     3241 gatttgcccc atcatgatga gaagacatgg aatgttggaa gcagcaaccg aaacaaagct
     3301 gaaaacctgt tgcgagggaa gcgagatggc acttttcttg tccgggagag cagtaaacag
     3361 ggctgctatg cctgctctgt agtggtggac ggcgaagtaa agcattgtgt cataaacaaa
     3421 acagcaactg gctatggctt tgccgagccc tataacttgt acagctctct gaaagaactg
     3481 gtgctacatt accaacacac ctcccttgtg cagcacaacg actccctcaa tgtcacacta
     3541 gcctacccag tatatgcaca gcagaggcga ctgaattctg cagatatcca gcacagtggc
     3601 ggccgctcga gagactacaa agaccatgac ggtgattata aagatcatga catcgactac
     3661 aaggatgacg atgacaagta gtctagaggg cccgatatct cgacaatcaa cctctggatt
     3721 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3781 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3841 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3901 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3961 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     4021 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     4081 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     4141 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     4201 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     4261 cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac
     4321 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4381 aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4441 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     4501 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     4561 gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta
     4621 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4681 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4741 ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc
     4801 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4861 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4921 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg
     4981 ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     5041 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     5101 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     5161 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     5221 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5281 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     5341 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     5401 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     5461 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     5521 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     5581 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     5641 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     5701 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     5761 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     5821 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5881 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5941 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     6001 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     6061 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     6121 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     6181 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     6241 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6301 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6361 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6421 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6481 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6541 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6601 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6661 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6721 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6781 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6841 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6901 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6961 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7021 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7081 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7141 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     7201 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7261 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7321 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7381 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     7441 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7501 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7561 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7621 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7681 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7741 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7801 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7861 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7921 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7981 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8041 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8101 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8161 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8221 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8281 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8341 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8401 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8461 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8521 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8581 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8641 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8701 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     8761 tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca
     8821 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     8881 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
