LOCUS       V043118                 7315 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043118
VERSION     V043118
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             937..1653
                     /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             1690..3066
                     /label="ZBTB12(NM_181842)"
                     /note="ZBTB12(NM_181842)"
                     /gene="ZBTB12"
     CDS             3079..3144
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3179..3403
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3449..3877
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3891..4220
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4232..4831
                     /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    4991..5112
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5161..5177)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5185..5201
                     /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(5209..5239)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5254..5275
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5563..6148)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6319..7179)
                     /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(7180..7284)
                     /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 aagctttcta gagaattcga tatcggatcc gccaccatgg tgagcaaggg cgaggagctg
      961 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1021 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1081 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1141 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1201 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1261 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1321 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1381 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1441 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1501 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1561 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1621 gggatcactc tcggcatgga cgagctgtac aagggcggtg gaggcagtgg tggaggcggt
     1681 agcggtacca tggcctctgg ggtggaagtc ctgcgcttcc agctgcccgg ccacgaggcc
     1741 gcaacgctac ggaacatgaa ccagctccgg gcagaggagc ggttctgcga cgtgaccatt
     1801 gtggccgaca gcctcaagtt tcgaggccac aaggtcatct tggccgcctg ctcacccttc
     1861 ctgcgggacc agttcctgct gaaccccagc tcggagctgc aggtctccct gatgcacagt
     1921 gcacgcatcg tggccgactt gctcctctcc tgctacacgg gcgccttgga attcgctgtt
     1981 agggacatcg tcaactacct tacagccgcc tcctacctgc agatggagca cgtggtggag
     2041 aaatgccgga atgccctcag ccagttcatt gagcccaaaa taggcctcaa agaggatggg
     2101 gtcagtgagg ctagccttgt gagcagcatc agcgccacca agtccctcct ccctccagcc
     2161 aggaccccaa agccagcccc gaagccccca cccccacctc ctctaccccc tccactcctg
     2221 cggccagtga agctggagtt cccactggat gaagacttgg agctgaaagc cgaggaagag
     2281 gatgaggatg aggatgagga cgtgtctgac atctgcatcg tcaaggtgga gtcggccctg
     2341 gaggtggcac accggctcaa accccctgga ggcctgggag ggggtctggg cattggaggc
     2401 tccgtgggtg gccaccttgg ggagctggcc cagagcagcg ttccccccag cactgtagcc
     2461 ccaccgcagg gtgtggtgaa ggcctgctat agcctgtcgg aagatgcaga aggggagggc
     2521 ctgctgttga ttcccggagg ccgggccagc gtgggggcca cctcgggcct ggtggaagca
     2581 gcagcggtgg ccatggctgc ccggggggcg gggggcagcc tgggggcggg gggcagccgg
     2641 ggacccctgc ctgggggctt ctcaggtgga aaccccttaa agaacatcaa gtgcaccaag
     2701 tgcccggaag tgttccaggg cgtggagaag ctggtcttcc acatgcgggc gcagcacttc
     2761 atcttcatgt gccctcgctg tggcaagcag ttcaaccaca gcagcaacct caaccgccac
     2821 atgaacgtgc atcgtggtgt caagtcacac tcgtgcggca tctgcggcaa gtgcttcaca
     2881 cagaagtcca cccttcacga ccacctcaac ctgcactcgg gagcgcggcc ctaccgctgc
     2941 tcctactgcg acgtgcgctt cgcccacaag cctgccatta ggcggcacct caaggagcaa
     3001 cacggcaaga ccacggccga gaacgtgctg gaggccagtg tggccgagat taacgtcctc
     3061 atccgcctcg agaccggtga ctacaaagac catgacggtg attataaaga tcatgacatc
     3121 gattacaagg atgacgatga caagtgaccg gtaagtttaa accgctgatc agcctcgact
     3181 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3241 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3301 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3361 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3421 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3481 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3541 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3601 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3661 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3721 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3781 cctatctcgg tctattcttt tgatttataa gggattttgg ccatttcggc ctattggtta
     3841 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3901 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3961 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4021 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4081 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4141 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4201 gaggcctagg cttttgcaaa aagctcccgg gatgaccgag tacaagccca cggtgcgcct
     4261 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     4321 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     4381 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4441 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4501 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     4561 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     4621 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     4681 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     4741 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     4801 ctggtgcatg acccgcaagc ccggtgcctg attcgaaatg accgaccaag cgacgcccaa
     4861 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4921 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4981 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5041 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5101 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5161 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5221 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5281 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5341 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5401 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5461 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5521 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5581 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5641 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5701 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5761 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5821 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5881 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5941 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6001 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6061 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     6121 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6181 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6241 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6301 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6361 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6421 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6481 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6541 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6601 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6661 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6721 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6781 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6841 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6901 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6961 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7021 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7081 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7141 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7201 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7261 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
