LOCUS       V025467                 7505 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025467
VERSION     V025467
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             941..2995
                     /label="DLL4(NM_019074)"
                     /note="DLL4(NM_019074)"
                     /gene="DLL4"
     CDS             3002..3067
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3105..3329
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3375..3803
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3817..4146
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4213..5007
                     /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    5181..5302
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5351..5367)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5375..5391
                     /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(5399..5429)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5444..5465
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5753..6338)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6509..7369)
                     /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(7370..7474)
                     /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 aagcttggta ccgagctcgg atccgccacc atggcggcag cgtcccggag
      961 cgcctctggc tgggcgctac tgctgctggt ggcactttgg cagcagcgcg cggccggctc
     1021 cggcgtcttc cagctgcagc tgcaggagtt catcaacgag cgcggcgtac tggccagtgg
     1081 gcggccttgc gagcccggct gccggacttt cttccgcgtc tgccttaagc acttccaggc
     1141 ggtcgtctcg cccggaccct gcaccttcgg gaccgtctcc acgccggtat tgggcaccaa
     1201 ctccttcgct gtccgggacg acagtagcgg cggggggcgc aaccctctcc aactgccctt
     1261 caatttcacc tggccgggta ccttctcgct catcatcgaa gcttggcacg cgccaggaga
     1321 cgacctgcgg ccagaggcct tgccaccaga tgcactcatc agcaagatcg ccatccaggg
     1381 ctccctagct gtgggtcaga actggttatt ggatgagcaa accagcaccc tcacaaggct
     1441 gcgctactct taccgggtca tctgcagtga caactactat ggagacaact gctcccgcct
     1501 gtgcaagaag cgcaatgacc acttcggcca ctatgtgtgc cagccagatg gcaacttgtc
     1561 ctgcctgccc ggttggactg gggaatattg ccaacagcct atctgtcttt cgggctgtca
     1621 tgaacagaat ggctactgca gcaagccagc agagtgcctc tgccgcccag gctggcaggg
     1681 ccggctgtgt aacgaatgca tcccccacaa tggctgtcgc cacggcacct gcagcactcc
     1741 ctggcaatgt acttgtgatg agggctgggg aggcctgttt tgtgaccaag atctcaacta
     1801 ctgcacccac cactccccat gcaagaatgg ggcaacgtgc tccaacagtg ggcagcgaag
     1861 ctacacctgc acctgtcgcc caggctacac tggtgtggac tgtgagctgg agctcagcga
     1921 gtgtgacagc aacccctgtc gcaatggagg cagctgtaag gaccaggagg atggctacca
     1981 ctgcctgtgt cctccgggct actatggcct gcattgtgaa cacagcacct tgagctgcgc
     2041 cgactccccc tgcttcaatg ggggctcctg ccgggagcgc aaccaggggg ccaactatgc
     2101 ttgtgaatgt ccccccaact tcaccggctc caactgcgag aagaaagtgg acaggtgcac
     2161 cagcaacccc tgtgccaacg ggggacagtg cctgaaccga ggtccaagcc gcatgtgccg
     2221 ctgccgtcct ggattcacgg gcacctactg tgaactccac gtcagcgact gtgcccgtaa
     2281 cccttgcgcc cacggtggca cttgccatga cctggagaat gggctcatgt gcacctgccc
     2341 tgccggcttc tctggccgac gctgtgaggt gcggacatcc atcgatgcct gtgcctcgag
     2401 tccctgcttc aacagggcca cctgctacac cgacctctcc acagacacct ttgtgtgcaa
     2461 ctgcccttat ggctttgtgg gcagccgctg cgagttcccc gtgggcttgc cgcccagctt
     2521 cccctgggtg gccgtctcgc tgggtgtggg gctggcagtg ctgctggtac tgctgggcat
     2581 ggtggcagtg gctgtgcggc agctgcggct tcgacggccg gacgacggca gcagggaagc
     2641 catgaacaac ttgtcggact tccagaagga caacctgatt cctgccgccc agcttaaaaa
     2701 cacaaaccag aagaaggagc tggaagtgga ctgtggcctg gacaagtcca actgtggcaa
     2761 acagcaaaac cacacattgg actataatct ggccccaggg cccctggggc gggggaccat
     2821 gccaggaaag tttccccaca gtgacaagag cttaggagag aaggcgccac tgcggttaca
     2881 cagtgaaaag ccagagtgtc ggatatcagc gatatgctcc cccagggact ccatgtacca
     2941 gtctgtgtgt ttgatatcag aggagaggaa tgaatgtgtc attgccacgg aggtactcga
     3001 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     3061 tgacaagtag tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     3121 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3181 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3241 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3301 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3361 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3421 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3481 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3541 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3601 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3661 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3721 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3781 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3841 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3901 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3961 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4021 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4081 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4141 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4201 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4261 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4321 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4381 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4441 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4501 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4561 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4621 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4681 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4741 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4801 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4861 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4921 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4981 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5041 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5101 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5161 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5221 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5281 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5341 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5401 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5461 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5521 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5581 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5641 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5701 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5761 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5821 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5881 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5941 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6001 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6061 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6121 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6181 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6241 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     6301 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6361 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6421 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6481 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6541 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6601 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6661 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6721 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6781 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6841 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6901 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6961 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7021 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7081 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7141 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7201 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7261 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7321 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7381 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7441 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7501 acgtc
//
