LOCUS       V022225                 8090 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022225
VERSION     V022225
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..2737
                     /label="Dll4(NM_001107760)"
                     /note="Dll4(NM_001107760)"
                     /gene="Dll4"
     CDS             2777..2842
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2866..3454
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3520..3568
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3770..4198
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4212..4541
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4608..5402
                     /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    5578..5699
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5748..5764)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5772..5788
                     /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(5796..5826)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5841..5862
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6150..6738)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6909..7769)
                     /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(7770..7874)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgacgcc tgggtcccgg agcgcctgtc gctgggcgct
      721 actgctgctg gcggtcctgt ggccgcagcg cgcggcgggc tccggcatct tccagttgcg
      781 gctgcaggag ttcgccaacg agcgcggtat gctggccaat gggcggccct gcgagcccgg
      841 ctgccggact ttcttccgaa tctgccttaa gcattaccag gcaaccttct ccgagggacc
      901 ctgcaccttt ggcaatgtct ccacgccggt attgggcacc aactccttcg tcatcaggga
      961 caagaatagc ggcagtggtc gtaaccctct tcaattgccc ttcaatttta cctggccggg
     1021 aaccttctca ctcaacatcc aagcttggca cacgccggga gacgacctgc ggccagagac
     1081 ttcaccagga aattcactta tcagccaaat catcatccaa ggctcccttg ctgtgggtaa
     1141 gaattggaaa tcagatgagc aaaataacac cctcacaaga ctgcgctact cttaccgggt
     1201 cgtctgcagc gacaactact atggagatag ctgttctcgc ctgtgcaaga agcgcgatga
     1261 ccacttcgga cattatgagt gccaaccaga tggcagcctg tcctgcctgc cgggctggac
     1321 tgggaaatac tgtgatcagc ctatatgtct ttctggctgt catgagcaga atggctactg
     1381 cagcaagcca gatgagtgca actgccgtcc aggttggcag ggccccttat gcaatgaatg
     1441 tatcccccac aatggctgtc gtcatggcac ctgcaccatc ccctggcaat gtgcctgcga
     1501 tgagggctgg ggaggtctgt tttgtgacca agatcttaat tactgcactc accactctcc
     1561 gtgcaagaat ggatcaactt gctccaacag tgggccgagg ggctatacct gcacctgtct
     1621 cccaggctac actggtgagc actgtgagct ggaactcagc aagtgtgcca gcaacccctg
     1681 tcgaaatggt ggcagctgta aggaccatga gaatagctac cactgcctgt gtcccccggg
     1741 ctactatggc cagcactgtg aacacagtac cttgacctgt gcggattcac cctgcttcaa
     1801 tggaggctcc tgccgggagc gcaaccaggg ggccagttat gcctgcgagt gtccccctaa
     1861 cttcactggg tccaactgtg agaagaaagt tgacaggtgt accagcaacc cctgtgccaa
     1921 tggaggtcag tgcctgaaca gaggtccaag ccggacctgc cgctgtcgac ctggattcac
     1981 aggcacccac tgtgaactgc acatcagcga ttgtgcccga agcccttgtg cccatggggg
     2041 cacttgccac gatctggaga acgggcctgt gtgcacctgc cctgctggct tttctggcag
     2101 gcgctgcgag gtgcggataa ccaacgacgc ctgtgcctca ggaccctgct tcaatggggc
     2161 cacttgctac actggcctct ccccaaacaa ctttgtctgc aactgtcctt atggctttgt
     2221 gggcagccgc tgcgagtttc ccgtgggctt gccacccagc ttcccctggg tagctgtctc
     2281 cctgggtgtg gggctggtgg tactgctggt gttgctggtc atggtggcag tggctgtgcg
     2341 gcagctgcgg cttcggaggc ccgatgacga cagcagggaa gccatgaaca atttgtcaga
     2401 cttccagaag gacaacctaa tccctgctgc ccagctcaag aacacaaacc agaagaagga
     2461 gctggaagtg gattgtggcc tggacaagtc caactgtggc aaactgcaga accacacact
     2521 ggactataat ctggccccag gattcctggg gcgggggagc acgcctggga agtatcctca
     2581 cagcgacaag agtttaggag agaaggtgcc acttcggtta cacagtgaaa agccggcatg
     2641 tcgaatatca gcgatatgct cccccaggga ctctatgtac cagtcagtat gtttgatatc
     2701 agaagagagg aacgaatgtg tcattgccac agaggtagaa ttctgcagat atccagcaca
     2761 gtggcggccg ctcgaggact acaaagacca tgacggtgat tataaagatc atgacatcga
     2821 ctacaaggat gacgatgaca agtagtgagg gcccgatatc tcgacaatca acctctggat
     2881 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2941 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3001 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3061 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3121 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3181 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3241 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     3301 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3361 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3421 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg ctaactgaaa
     3481 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     3541 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     3601 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     3661 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     3721 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
     3781 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3841 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3901 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     3961 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4021 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4081 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4141 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     4201 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4261 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4381 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4441 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4501 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4561 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4621 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4681 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4741 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4801 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4861 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4921 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4981 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5041 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5101 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5161 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5221 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5281 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5341 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5401 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5461 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5521 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5581 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5641 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5701 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5761 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5821 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5881 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5941 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6001 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6061 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6121 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6181 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6241 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6301 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     6361 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6421 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6481 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6541 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6601 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6661 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6721 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6781 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6841 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6901 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6961 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7021 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7081 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7141 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7201 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7261 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7321 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7381 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7441 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7501 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7561 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7621 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7681 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7741 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7801 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7861 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     7921 ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc
     7981 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     8041 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
