LOCUS       V018620                 9689 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018620
VERSION     V018620
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..4336
                     /label="JAG1(NM_000214)"
                     /note="JAG1(NM_000214)"
                     /gene="JAG1"
     CDS             4376..4441
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4465..5053
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5119..5167
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5369..5797
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5811..6140
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6207..7001
                     /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    7177..7298
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7347..7363)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7371..7387
                     /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(7395..7425)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7440..7461
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7749..8337)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8508..9368)
                     /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(9369..9473)
                     /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 ccatgcgttc cccacggacg cgcggccggt ccgggcgccc
      721 cctaagcctc ctgctcgccc tgctctgtgc cctgcgagcc aaggtgtgtg gggcctcggg
      781 tcagttcgag ttggagatcc tgtccatgca gaacgtgaac ggggagctgc agaacgggaa
      841 ctgctgcggc ggcgcccgga acccgggaga ccgcaagtgc acccgcgacg agtgtgacac
      901 atacttcaaa gtgtgcctca aggagtatca gtcccgcgtc acggccgggg ggccctgcag
      961 cttcggctca gggtccacgc ctgtcatcgg gggcaacacc ttcaacctca aggccagccg
     1021 cggcaacgac cgcaaccgca tcgtgctgcc tttcagtttc gcctggccga ggtcctatac
     1081 gttgcttgtg gaggcgtggg attccagtaa tgacaccgtt caacctgaca gtattattga
     1141 aaaggcttct cactcgggca tgatcaaccc cagccggcag tggcagacgc tgaagcagaa
     1201 cacgggcgtt gcccactttg agtatcagat ccgcgtgacc tgtgatgact actactatgg
     1261 ctttggctgc aataagttct gccgccccag agatgacttc tttggacact atgcctgtga
     1321 ccagaatggc aacaaaactt gcatggaagg ctggatgggc cccgaatgta acagagctat
     1381 ttgccgacaa ggctgcagtc ctaagcatgg gtcttgcaaa ctcccaggtg actgcaggtg
     1441 ccagtacggc tggcaaggcc tgtactgtga taagtgcatc ccacacccgg gatgcgtcca
     1501 cggcatctgt aatgagccct ggcagtgcct ctgtgagacc aactggggcg gccagctctg
     1561 tgacaaagat ctcaattact gtgggactca tcagccgtgt ctcaacgggg gaacttgtag
     1621 caacacaggc cctgacaaat atcagtgttc ctgccctgag gggtattcag gacccaactg
     1681 tgaaattgct gagcacgcct gcctctctga tccctgtcac aacagaggca gctgtaagga
     1741 gacctccctg ggctttgagt gtgagtgttc cccaggctgg accggcccca catgctctac
     1801 aaacattgat gactgttctc ctaataactg ttcccacggg ggcacctgcc aggacctggt
     1861 taacggattt aagtgtgtgt gccccccaca gtggactggg aaaacgtgcc agttagatgc
     1921 aaatgaatgt gaggccaaac cttgtgtaaa cgccaaatcc tgtaagaatc tcattgccag
     1981 ctactactgc gactgtcttc ccggctggat gggtcagaat tgtgacataa atattaatga
     2041 ctgccttggc cagtgtcaga atgacgcctc ctgtcgggat ttggttaatg gttatcgctg
     2101 tatctgtcca cctggctatg caggcgatca ctgtgagaga gacatcgatg aatgtgccag
     2161 caacccctgt ttgaatgggg gtcactgtca gaatgaaatc aacagattcc agtgtctgtg
     2221 tcccactggt ttctctggaa acctctgtca gctggacatc gattattgtg agcctaatcc
     2281 ctgccagaac ggtgcccagt gctacaaccg tgccagtgac tatttctgca agtgccccga
     2341 ggactatgag ggcaagaact gctcacacct gaaagaccac tgccgcacga ccccctgtga
     2401 agtgattgac agctgcacag tggccatggc ttccaacgac acacctgaag gggtgcggta
     2461 tatttcctcc aacgtctgtg gtcctcacgg gaagtgcaag agtcagtcgg gaggcaaatt
     2521 cacctgtgac tgtaacaaag gcttcacggg aacatactgc catgaaaata ttaatgactg
     2581 tgagagcaac ccttgtagaa acggtggcac ttgcatcgat ggtgtcaact cctacaagtg
     2641 catctgtagt gacggctggg agggggccta ctgtgaaacc aatattaatg actgcagcca
     2701 gaacccctgc cacaatgggg gcacgtgtcg cgacctggtc aatgacttct actgtgactg
     2761 taaaaatggg tggaaaggaa agacctgcca ctcacgtgac agtcagtgtg atgaggccac
     2821 gtgcaacaac ggtggcacct gctatgatga gggggatgct tttaagtgca tgtgtcctgg
     2881 cggctgggaa ggaacaacct gtaacatagc ccgaaacagt agctgcctgc ccaacccctg
     2941 ccataatggg ggcacatgtg tggtcaacgg cgagtccttt acgtgcgtct gcaaggaagg
     3001 ctgggagggg cccatctgtg ctcagaatac caatgactgc agccctcatc cctgttacaa
     3061 cagcggcacc tgtgtggatg gagacaactg gtaccggtgc gaatgtgccc cgggttttgc
     3121 tgggcccgac tgcagaataa acatcaatga atgccagtct tcaccttgtg cctttggagc
     3181 gacctgtgtg gatgagatca atggctaccg gtgtgtctgc cctccagggc acagtggtgc
     3241 caagtgccag gaagtttcag ggagaccttg catcaccatg gggagtgtga taccagatgg
     3301 ggccaaatgg gatgatgact gtaatacctg ccagtgcctg aatggacgga tcgcctgctc
     3361 aaaggtctgg tgtggccctc gaccttgcct gctccacaaa gggcacagcg agtgccccag
     3421 cgggcagagc tgcatcccca tcctggacga ccagtgcttc gtccacccct gcactggtgt
     3481 gggcgagtgt cggtcttcca gtctccagcc ggtgaagaca aagtgcacct ctgactccta
     3541 ttaccaggat aactgtgcga acatcacatt tacctttaac aaggagatga tgtcaccagg
     3601 tcttactacg gagcacattt gcagtgaatt gaggaatttg aatattttga agaatgtttc
     3661 cgctgaatat tcaatctaca tcgcttgcga gccttcccct tcagcgaaca atgaaataca
     3721 tgtggccatt tctgctgaag atatacggga tgatgggaac ccgatcaagg aaatcactga
     3781 caaaataatc gatcttgtta gtaaacgtga tggaaacagc tcgctgattg ctgccgttgc
     3841 agaagtaaga gttcagaggc ggcctctgaa gaacagaaca gatttccttg ttcccttgct
     3901 gagctctgtc ttaactgtgg cttggatctg ttgcttggtg acggccttct actggtgcct
     3961 gcggaagcgg cggaagccgg gcagccacac acactcagcc tctgaggaca acaccaccaa
     4021 caacgtgcgg gagcagctga accagatcaa aaaccccatt gagaaacatg gggccaacac
     4081 ggtccccatc aaggattatg agaacaagaa ctccaaaatg tctaaaataa ggacacacaa
     4141 ttctgaagta gaagaggacg acatggacaa acaccagcag aaagcccggt ttgccaagca
     4201 gccggcgtac acgctggtag acagagaaga gaagcccccc aacggcacgc cgacaaaaca
     4261 cccaaactgg acaaacaaac aggacaacag agacttggaa agtgcccaga gcttaaaccg
     4321 aatggagtac atcgtagaat tctgcagata tccagcacag tggcggccgc tcgaggacta
     4381 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     4441 gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     4501 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4561 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4621 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4681 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4741 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4801 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
     4861 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4921 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4981 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     5041 gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     5101 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     5161 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5221 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5281 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     5341 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     5401 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5461 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5521 cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5581 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5641 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5701 cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta
     5761 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     5821 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5881 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5941 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     6001 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     6061 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     6121 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     6181 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     6241 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     6301 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     6361 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     6421 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     6481 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     6541 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     6601 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     6661 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     6721 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     6781 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     6841 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6901 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6961 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     7021 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     7081 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     7141 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     7201 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     7261 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     7321 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     7381 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     7441 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     7501 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7561 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7621 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7681 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7741 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7801 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7861 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7921 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     7981 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     8041 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     8101 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     8161 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     8221 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     8281 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     8341 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     8401 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     8461 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     8521 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     8581 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     8641 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     8701 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8761 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8821 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8881 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8941 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     9001 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     9061 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     9121 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     9181 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     9241 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     9301 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     9361 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     9421 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     9481 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga
     9541 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     9601 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     9661 ccgacaattg catgaagaat ctgcttagg
//
