LOCUS       V037404                10355 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037404
VERSION     V037404
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             923..1639
                     /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             1676..5845
                     /label="ATAD2(NM_014109)"
                     /note="ATAD2(NM_014109)"
                     /gene="ATAD2"
     CDS             5852..5917
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5955..6179
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6225..6653
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6667..6996
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7063..7857
                     /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    8031..8152
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8201..8217)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8225..8241
                     /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(8249..8279)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8294..8315
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8603..9188)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9359..10219)
                     /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(10220..10324)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      961 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1021 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1081 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     1141 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1201 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1261 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1321 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1381 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1441 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1501 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1561 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1621 catggacgag ctgtacaagg gcggtggagg cagtggtgga ggcggtagcg aattcatggt
     1681 ggttctccgc agcagcttgg agctgcacaa ccactccgcg gcctcggcca cgggctcctt
     1741 ggacctgtcc agtgacttcc tcagtctgga gcacatcggc cggaggcggc tccgctcggc
     1801 cggcgcggcg cagaagaaac ccgcggcgac cacagccaaa gcgggcgatg ggtcatcagt
     1861 taaggaagtt gaaacctacc accggacacg tgctttaaga tctttgagaa aagatgcaca
     1921 gaattcttca gattctagtt ttgagaagaa tgtggaaata acggagcaac ttgctaatgg
     1981 caggcatttt acaaggcagt tggccagaca gcaggctgat aaaaaaaaag aagagcacag
     2041 agaagacaaa gtgattccag ttactcggtc attgagggct agaaacatcg ttcaaagtac
     2101 agaacactta catgaagata atggtgatgt tgaagtgcgt cgaagttgta ggattagaag
     2161 tcgttatagt ggtgtaaacc agtccatgct gtttgacaaa cttataacta acactgctga
     2221 agctgtactt caaaaaatgg atgacatgaa gaagatgcgt agacagcgaa tgagagaact
     2281 tgaagacttg ggagtgttta atgaaacaga agaaagcaat cttaatatgt acacaagagg
     2341 aaaacagaaa gatattcaaa gaactgatga agaaacaact gataatcaag aaggcagtgt
     2401 ggagtcatct gaagagggtg aagaccaaga acatgaagat gatggtgaag atgaagatga
     2461 tgaagatgat gatgatgatg acgatgatga tgatgatgat gatgatgaag atgatgaaga
     2521 tgaagaagat ggagaagaag agaatcagaa gcgatattat cttagacaga gaaaagctac
     2581 tgtttactat caggctccat tggaaaaacc tcgtcaccag agaaagccca acatatttta
     2641 tagtggccca gcttctcctg caagaccaag ataccgatta tcttccgcag gaccaagaag
     2701 tccttactgt aaacgaatga acaggcgaag gcatgcaatc cacagtagtg actcgacttc
     2761 atcttcctcc tctgaagatg aacagcactt tgagaggcgg aggaaaagga gtcgtaatag
     2821 ggctatcaat aggtgcctcc cactaaattt tcggaaagat gaattaaaag gcatttataa
     2881 agatcgaatg aaaattggag caagccttgc cgatgttgat ccaatgcaac tagattcttc
     2941 agtacgattt gatagtgttg gtggcctgtc taatcatata gcagctctaa aagagatggt
     3001 ggtgtttcca ttactttatc cagaagtctt tgaaaaattt aaaattcaac ccccaagagg
     3061 ttgtttgttt tatgggccac ctggaactgg aaagactctg gttgccagag cacttgccaa
     3121 tgagtgcagt caaggggata aaagagtagc atttttcatg aggaaaggtg ctgattgtct
     3181 aagtaaatgg gtaggagaat ctgaaagaca gctacgattg ctgtttgatc aggcctatca
     3241 gatgcgccca tcaattattt tttttgacga aattgatggt ctggctccag tacggtcaag
     3301 caggcaagat cagattcaca gttctattgt ttccaccctg ctagctctta tggatggatt
     3361 ggacagcaga ggggaaattg tggtcattgg tgctacgaac aggctagatt ctatagatcc
     3421 tgctttacga aggcctggtc gctttgatag agaattcctc tttagcctgc ctgataaaga
     3481 ggctcgaaaa gagattctaa agattcacac cagggattgg aatcccaaac cactggacac
     3541 atttttagaa gagctagcag aaaactgtgt tggatactgt ggagcagata ttaaatcaat
     3601 atgtgctgaa gctgctttat gtgctttacg acgacgctac ccacagatct ataccactag
     3661 tgagaaactg cagttggatc tctcttcaat taatatctca gctaaggatt tcgaggtagc
     3721 tatgcaaaag atgataccag cctcccaaag agctgtgaca tcacctgggc aggcactgtc
     3781 caccgttgtg aaaccactcc tgcaaaacac tgttgacaag attttagaag ccctgcagag
     3841 agtatttcca catgcagaat tcagaacaaa taaaacatta gactcagata tttcttgtcc
     3901 tctgctagaa agtgacttgg cttacagtga tgatgatgtt ccatcagttt atgaaaatgg
     3961 actttctcag aaatcttctc ataaggcaaa agacaatttt aattttcttc atttgaatag
     4021 aaatgcttgt taccaaccta tgtcttttcg accaagaata ttgatagtag gagaaccagg
     4081 atttgggcaa ggttctcact tggcaccagc tgtcattcat gctttggaaa agtttactgt
     4141 atatacatta gacattcctg ttctttttgg agttagtact acatcccctg aagaaacatg
     4201 tgcccaggtg attcgtgaag ctaagagaac agcaccaagt atagtgtatg ttcctcatat
     4261 ccacgtgtgg tgggaaatag ttggaccgac acttaaagcc acatttacca cattattaca
     4321 gaatattcct tcatttgctc cagttttact acttgcaact tctgacaaac cccattccgc
     4381 tttgccagaa gaggtgcaag aattgtttat ccgtgattat ggagagattt ttaatgtcca
     4441 gttaccggat aaagaagaac ggacaaaatt ttttgaagat ttaattctaa aacaagctgc
     4501 taagcctcct atatcaaaaa agaaagcagt tttgcaggct ttggaggtac tcccagtagc
     4561 accaccacct gagccaagat cactgacagc agaagaagtg aaacgactag aagaacaaga
     4621 agaagataca tttagagaac tgaggatttt cttaagaaat gttacacata ggcttgctat
     4681 tgacaagcga ttccgagtgt ttactaagcc tgttgaccct gatgaggttc ctgattatgt
     4741 cactgtaata aagcaaccaa tggacctttc atctgtaatc agtaaaattg atctacacaa
     4801 gtatctgact gtgaaagact atttgagaga tattgatcta atctgtagta atgccttaga
     4861 atacaatcca gatagagatc ctggagatcg tcttattagg catagagcct gtgctttaag
     4921 agatactgcc tatgccataa ttaaagaaga acttgatgaa gactttgagc agctctgtga
     4981 agaaattcag gaatctagaa agaaaagagg ttgtagctcc tccaaatatg ccccgtctta
     5041 ctaccatgtg atgccaaagc aaaattccac tcttgttggt gataaaagat cagacccaga
     5101 gcagaatgaa aagctaaaga caccgagtac tcctgtggct tgcagcactc ctgctcagtt
     5161 gaagaggaaa attcgcaaaa agtcaaactg gtacttaggc accataaaaa agcgaaggaa
     5221 gatttcacag gcaaaggatg atagccagaa tgccatagat cacaaaattg agagtgatac
     5281 agaggaaact caagacacaa gtgtagatca taatgagacc ggaaacacag gagagtcttc
     5341 ggtggaagaa aatgaaaaac agcaaaatgc ctctgaaagc aaactggaat tgagaaataa
     5401 ttcaaatact tgtaatatag agaatgagct tgaagactct aggaagacta cagcatgtac
     5461 agaattgaga gacaagattg cttgtaatgg agatgcttct agctctcaga taatacatat
     5521 ttctgatgaa aatgaaggaa aagaaatgtg tgttctgcga atgactcgag ctagacgttc
     5581 ccaggtagaa cagcagcagc tcatcactgt tgaaaaggct ttggcaattc tttctcagcc
     5641 tacaccctca cttgttgtgg atcatgagcg attaaaaaat cttttgaaga ctgttgttaa
     5701 aaaaagtcaa aactacaaca tatttcagtt ggaaaatttg tatgcagtaa tcagccaatg
     5761 tatttatcgg catcgcaagg accatgataa aacatcactt attcagaaaa tggagcaaga
     5821 ggtagaaaac ttcagttgtt ccagactcga ggactacaaa gaccatgacg gtgattataa
     5881 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg
     5941 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     6001 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     6061 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     6121 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     6181 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     6241 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6301 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6361 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6421 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6481 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6541 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6601 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     6661 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     6721 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     6781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     6841 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     6901 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     6961 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     7021 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     7081 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     7141 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     7201 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     7261 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     7321 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     7381 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     7441 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     7501 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     7561 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     7621 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     7681 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     7741 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     7801 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     7861 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     7921 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     7981 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     8041 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     8101 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     8161 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     8221 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     8281 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     8341 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     8401 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     8461 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     8521 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     8581 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     8641 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8701 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8761 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8821 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8881 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8941 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     9001 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     9061 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     9121 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     9181 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     9241 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     9301 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     9361 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     9421 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     9481 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     9541 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     9601 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     9661 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     9721 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     9781 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     9841 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     9901 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     9961 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
    10021 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
    10081 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
    10141 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
    10201 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
    10261 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
    10321 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
