LOCUS       V028799                 7638 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028799
VERSION     V028799
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             1627..3249
                     /label="ATL3(NM_015459)"
                     /note="ATL3(NM_015459)"
                     /gene="ATL3"
     CDS             3256..3321
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3400..3624
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3670..4098
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4112..4441
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4508..5107
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5281..5402
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5451..5467)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5475..5491
                     /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(5499..5529)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5544..5565
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5853..6438)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6609..7469)
                     /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(7470..7574)
                     /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 aagcttggtg gaggcggatc aggaggcggt
     1621 ggatctatgt tgtcccctca gcgagtggca gcagctgcct caagaggagc agatgatgcc
     1681 atggagagca gcaagcctgg tccagtgcag gttgttttgg ttcagaaaga tcaacattcc
     1741 tttgagctag atgagaaagc cttggccagc atcctcttgc aggaccacat ccgagatctt
     1801 gatgtggtgg tggtttcagt ggctggtgcc ttccgaaagg gcaagtcctt cattctggat
     1861 tttatgctac gatacttata ttctcagaag gaaagtggcc attcaaattg gttgggtgac
     1921 ccagaagaac cgttaacagg attttcctgg agagggggat ctgatccaga aaccactggg
     1981 attcaaatct ggagtgaagt tttcactgtg gagaagccag gtgggaagaa ggttgcagtt
     2041 gttctgatgg atacccaggg ggcatttgac agccagtcaa ctgtgaaaga ctgtgctacc
     2101 atctttgctc taagcactat gactagttct gttcagattt ataatttatc tcagaacatt
     2161 caagaagatg atcttcaaca gctgcagctc ttcacagaat acggtcgtct ggcaatggat
     2221 gaaattttcc aaaagccttt ccagacactg atgtttttgg ttagagattg gagtttccct
     2281 tatgaatata gctatggact ccaaggagga atggcatttt tggataagcg tttacaggtg
     2341 aaggaacatc aacatgaaga aattcagaat gttcgaaatc acattcactc atgtttctcc
     2401 gatgtcacct gctttctctt accacatcca ggactccagg tggccacaag ccctgacttt
     2461 gatgggaaat taaaagatat tgctggtgaa ttcaaagagc agttacaggc actgataccg
     2521 tatgtattaa acccatctaa gttaatggaa aaggagatca atggctcaaa ggtcacctgt
     2581 cggggactac tggagtattt taaggcatat attaaaattt atcaaggaga agatctgcct
     2641 caccccaagt ccatgcttca ggccactgct gaagccaaca acttagcagc tgcagcctct
     2701 gccaaggaca tttattataa caacatggaa gaggtttgtg ggggagagaa accttatttg
     2761 tctccagaca ttctagagga gaagcactgt gaattcaaac aacttgctct ggaccatttt
     2821 aagaagacca agaagatggg tgggaaggat ttcagctttc gttaccagca ggagctggag
     2881 gaggaaatca aggaattata tgagaacttc tgcaagcaca atggtagcaa gaacgtcttc
     2941 agcaccttcc gaacccctgc agtgctgttc acgggcattg tagctttgta catagcctca
     3001 ggcctcactg gcttcatagg tcttgaggtt gtagcccagt tgttcaactg tatggttgga
     3061 ctactgttaa tagcactcct cacctggggc tacatcaggt attctggtca atatcgtgag
     3121 ctgggcggag ctattgattt tggtgccgca tatgtgttgg agcaggcttc ttctcatatc
     3181 ggtaattcca ctcaggccac tgtgagggat gcagttgttg gaagaccatc catggataaa
     3241 aaagctcaac tcgaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     3301 tacaaggatg acgatgacaa gtagaattct gcagatatcc agcacagtgg cggccgctcg
     3361 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3421 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3481 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3541 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3601 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3661 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3721 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3781 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3841 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3901 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3961 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4021 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4081 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4141 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4201 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4261 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4321 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4381 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4441 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4501 gtttcgcatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg
     4561 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     4621 cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     4681 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     4741 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     4801 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     4861 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     4921 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     4981 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     5041 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     5101 tgcctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5161 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5221 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5281 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5341 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5401 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5461 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5521 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5581 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5641 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5701 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5761 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5821 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5881 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5941 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6001 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6061 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6121 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6181 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6241 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6301 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6361 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc
     6421 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6481 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6541 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6601 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6661 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6721 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6781 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6841 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6901 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6961 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7021 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7081 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7141 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7201 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7261 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7321 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7381 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7441 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7501 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7561 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     7621 ctcccgatcc cctatggt
//
