LOCUS       V027254                 8751 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027254
VERSION     V027254
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /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             1642..4146
                     /label="TRIM28(NM_005762)"
                     /note="TRIM28(NM_005762)"
                     /gene="TRIM28"
     CDS             4153..4242
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    4318..4542
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4588..5016
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5030..5359
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5426..6220
                     /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    6394..6515
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6564..6580)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6588..6604
                     /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(6612..6642)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6657..6678
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6966..7551)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7722..8582)
                     /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(8583..8687)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catggcggcc tccgcggcgg cagcctcggc agcagcggcc
     1681 tcggccgcct ctggcagccc gggcccgggc gagggctccg ctggcggcga aaagcgctcc
     1741 accgcccctt cggccgcagc ctcggcctct gcctcagccg cggcgtcgtc gcccgcgggg
     1801 ggcggcgccg aggcgctgga gctgctggag cactgcggcg tgtgcagaga gcgcctgcga
     1861 cccgagaggg agccccgcct gctgccctgt ttgcactcgg cctgtagtgc ctgcttaggg
     1921 cccgcggccc ccgccgccgc caacagctcg ggggacggcg gggcggcggg cgacggcacc
     1981 gtggtggact gtcccgtgtg caagcaacag tgcttctcca aagacatcgt ggagaattat
     2041 ttcatgcgtg atagtggcag caaggctgcc accgacgccc aggatgcgaa ccagtgctgc
     2101 actagctgtg aggataatgc cccagccacc agctactgtg tggagtgctc ggagcctctg
     2161 tgtgagacct gtgtagaggc gcaccagcgg gtgaagtaca ccaaggacca tactgtgcgc
     2221 tctactgggc cagccaagtc tcgggatggt gaacgtactg tctattgcaa cgtacacaag
     2281 catgaacccc ttgtgctgtt ttgtgagagc tgtgatactc tcacctgccg agactgccag
     2341 ctcaatgccc acaaggacca ccagtaccag ttcttagagg atgcagtgag gaaccagcgc
     2401 aagctcctgg cctcactggt gaagcgcctt ggggacaaac atgcaacatt gcagaagagc
     2461 accaaggagg ttcgcagctc aatccgccag gtgtctgacg tacagaagcg tgtgcaagtg
     2521 gatgtcaaga tggccatcct gcagatcatg aaggagctga ataagcgggg ccgtgtgctg
     2581 gtcaatgatg cccagaaggt gactgagggg cagcaggagc gcctggagcg gcagcactgg
     2641 accatgacca agatccagaa gcaccaggag cacattctgc gctttgcctc ttgggctctg
     2701 gagagtgaca acaacacagc ccttttgctt tctaagaagt tgatctactt ccagctgcac
     2761 cgggccctca agatgattgt ggatcccgtg gagccacatg gcgagatgaa gtttcagtgg
     2821 gacctcaatg cctggaccaa gagtgccgag gcctttggca agattgtggc agagcgtcct
     2881 ggcactaact caacaggccc tgcacccatg gcccctccaa gagccccagg gcccctgagc
     2941 aagcagggct ctggcagcag ccagcccatg gaggtgcagg aaggctatgg ctttgggtca
     3001 ggagatgatc cctactcaag tgcagagccc catgtgtcag gtgtgaaacg gtcccgctca
     3061 ggtgagggcg aggtgagcgg ccttatgcgc aaggtgccac gagtgagcct tgaacgcctg
     3121 gacctggacc tcacagctga cagccagcca cccgtcttca aggtcttccc aggcagtacc
     3181 actgaggact acaaccttat tgttattgaa cgtggcgctg ccgctgcagc taccggccag
     3241 ccagggactg cgcctgcagg aacccctggt gccccacccc tggctggcat ggccattgtc
     3301 aaggaggagg agacggaggc tgccattgga gcccctccta ctgccactga gggccctgag
     3361 accaaacctg tgcttatggc tcttgcggag ggtcctggtg ctgagggtcc ccgcctggcc
     3421 tcacctagtg gcagcaccag ctcagggctg gaggtggtgg ctcctgaggg tacctcagcc
     3481 ccaggtggtg gcccgggaac cctggatgac agtgccacca tttgccgtgt ctgccagaag
     3541 ccaggcgatc tggttatgtg caaccagtgt gagttttgtt tccacctgga ctgtcacctg
     3601 ccggccctgc aggatgtacc aggggaggag tggagctgct cactctgcca tgtgctccct
     3661 gacctgaagg aggaggatgg cagcctcagc ctggatggtg cagacagcac tggcgtggtg
     3721 gccaagctct caccagccaa ccagcggaaa tgtgagcgtg tactgctggc cctattctgt
     3781 cacgaaccct gccgccccct gcatcagctg gctaccgact ccaccttctc cctggaccag
     3841 cccggtggca ccctggatct gaccctgatc cgtgcccgcc tccaggagaa gttgtcacct
     3901 ccctacagct ccccacagga gtttgcccag gatgtgggcc gcatgttcaa gcaattcaac
     3961 aagttaactg aggacaaggc agacgtgcag tccatcatcg gcctgcagcg cttcttcgag
     4021 acgcgcatga acgaggcctt cggtgacacc aagttctctg ctgtgctggt ggagcccccg
     4081 ccgatgagcc tgcctggtgc tggcctgagt tcccaggagc tgtctggtgg ccctggtgat
     4141 ggccccaagc ttgaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt
     4201 gaagaagatt tagagcaaaa gttaatttcc gaggaggact tataggatcc tgatatccag
     4261 cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     4321 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4381 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4441 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4501 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     4561 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     4621 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4681 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4741 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4801 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4861 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4921 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4981 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     5041 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5101 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5161 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     5221 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     5281 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     5341 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     5401 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     5461 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5521 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5581 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     5641 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5701 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     5761 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5821 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5881 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5941 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     6001 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6061 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6121 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6181 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     6241 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     6301 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     6361 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     6421 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     6481 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     6541 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     6601 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     6661 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6721 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6781 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6841 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6901 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6961 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     7021 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7081 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7141 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7201 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7261 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7321 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7381 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7441 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7501 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7561 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7621 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7681 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7741 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7801 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     7861 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     7921 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     7981 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     8041 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8101 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8161 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8221 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8281 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     8341 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8401 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8461 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8521 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8581 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8641 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8701 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg t
//
