LOCUS       V022146                 9692 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022146
VERSION     V022146
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..4339
                     /label="Jag1(NM_019147)"
                     /note="Jag1(NM_019147)"
                     /gene="Jag1"
     CDS             4379..4444
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4468..5056
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5122..5170
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5372..5800
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5814..6143
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6210..7004
                     /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    7180..7301
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7350..7366)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7374..7390
                     /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(7398..7428)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7443..7464
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7752..8340)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8511..9371)
                     /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(9372..9476)
                     /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 ccatgcggtc cccacggacg cgcggccggc cggggcgccc
      721 cttgagcctt ctgctcgccc tgctctgtgc cctgcgagcc aaggtgtgcg gggcctcggg
      781 tcaatttgag ctggagatcc tgtccatgca gaacgtgaac ggagagttac agaatgggaa
      841 ctgttgcggc ggagcccgga accctggcga ccgcaagtgc acccgcgacg agtgtgacac
      901 gtacttcaaa gtgtgcctca aggagtatca gtcccgcgtc acggccgggg gaccctgcag
      961 cttcggctca gggtctacgc ctgtcatcgg gggcaatacc ttcaatctca aggccagccg
     1021 cggcaacgac cgtaatcgca tcgtactgcc tttcagtttc gcctggccga ggtcctacac
     1081 tttgctggtg gaggcatggg attccagtaa tgacactatt caacctgata gcataattga
     1141 aaaggcttct cactcaggca tgataaaccc cagccggcag tggcagacac tgaaacagaa
     1201 cacagggatt gcccactttg agtatcagat ccgagtgacc tgtgatgacc actactatgg
     1261 ctttggctgc aacaagttct gtcggcccag agatgacttc tttggacatt atgcctgcga
     1321 ccagaatggc aacaaaacct gcatggaagg ctggatgggt cctgagtgta acaaagcaat
     1381 atgccgacag ggctgcagtc ccaagcatgg ctcctgcaaa ctcccaggtg actgcaggtg
     1441 ccagtatggt tggcagggcc tgtactgtga taagtgcatc ccgcacccag gatgtgtcca
     1501 cggcacctgc aatgaaccct ggcagtgcct ctgtgaaacc aactggggtg gacagctctg
     1561 tgacaaagat ctgaattact gtgggactca tcagccctgt ctcaaccggg gaacatgtag
     1621 caacactggc cctgacaaat accagtgttc ctgccccgag ggctactcag gaccgaactg
     1681 tgaaattgct gagcatgcct gcctctctga tccctgccac aacagaggca gctgcaagga
     1741 gacctcctcg ggctttgagt gtgagtgttc tccaggctgg actggaccca cgtgttccac
     1801 aaacatcgat gactgttctc caaataactg ttcccatgga ggcacctgcc aggatctagt
     1861 gaatggattc aagtgcgtgt gcccacccca gtggactggc aagacttgtc agttagatgc
     1921 aaatgagtgc gaggccaagc cttgtgtaaa tgccagatcc tgtaagaatc tgattgccag
     1981 ctactactgt gattgtcttc ctggctggat gggtcagaat tgtgacataa atatcaatga
     2041 ctgccttggc cagtgtcaga atgatgcctc ctgtcgggat ttggttaatg gttatcgctg
     2101 tatctgtcca cctggctatg cgggcgatca ctgtgaaaga gacatcgatg aatgtgctag
     2161 caacccctgc ttgaatgggg gtcactgtca gaatgaaatt aacagattcc agtgtctgtg
     2221 tcccactggt ttctctggaa acctctgtca gctggacatc gattactgcg agcccaaccc
     2281 ttgccagaat ggcgcccaat gctacaatcg tgccagtgac tatttctgca agtgccctga
     2341 ggactacgag ggcaagaact gctcacacct gaaagaccac tgccgcacca ctccctgtga
     2401 agtgattgac agctgcaccg tggccatggc ctccaacgat acgcctgaag gggtgcggta
     2461 tatctcttcc aacgtctgtg gtccccatgg gaagtgcaag agcgagtcag gaggcaaatt
     2521 cacctgtgac tgtaacaaag gcttcaccgg cacctactgc catgaaaata tcaacgactg
     2581 tgagggcaac ccctgtacga acggtggcac gtgcatcgac ggcgtcaact cctacaagtg
     2641 tatctgtagt gacggctggg agggcgcgca ctgtgagaac aacataaatg actgtagcca
     2701 gaacccttgc cactatgggg gtacatgtcg agacctggtc aatgactttt actgtgactg
     2761 caaaaatggg tggaaaggga aaacctgcca ctcgcgtgac agccagtgtg acgaagccac
     2821 gtgtaataac ggtggtacct gctatgatga agtggatacg tttaagtgca tgtgtcctgg
     2881 tggctgggaa ggaacaacct gtaatatagc tagaaacagt agctgcctgc cgaacccctg
     2941 tcataatgga ggcacctgcg tggtcaatgg agactccttc acctgtgtct gcaaagaagg
     3001 ctgggagggg ccgatttgta ctcaaaatac caacgactgc agtcctcacc cttgttacaa
     3061 cagcggaacc tgtgtggatg gagataactg gtaccggtgc gaatgtgccc ctggttttgc
     3121 tgggccagac tgcaggataa acatcaatga gtgtcagtct tccccttgtg cctttggggc
     3181 cacctgtgtg gatgagatca atggctacca gtgtatctgc cctccaggac atagtggtgc
     3241 caagtgccat gaagtttcag ggagatcttg catcaccatg gggagagtga tacttgatgg
     3301 ggccaagtgg gatgacgact gtaacacctg ccagtgcctg aatggacggg tggcctgctc
     3361 caaggtctgg tgtggcccga gaccttgcca gctccacaaa gggcacggcg agtgtcccaa
     3421 tgggcagagc tgcatcccag tcctggatga ccagtgtttc gtgcgcccct gcactggtgc
     3481 tggcgagtgt cggtcctcca gcctccagcc agtgaagacc aagtgtacat cggactctta
     3541 ttaccaggac aattgtgcga acatcacttt cacctttaac aaggagatga tgtctccagg
     3601 tctcaccact gagcacattt gcagcgaatt gaggaatctg aatattctga agaatgtttc
     3661 tgctgagtat tcaatctaca tagcctgtga gccttcctta tcagcaaaca atgaaataca
     3721 cgtggccatt tctgccgaag acatccggga tgatggaaac cctgtcaagg aaattactga
     3781 taaaataata gatctcgtta gtaaacggga tgggaacagc tcacttatcg ctgctgttgc
     3841 agaagtcaga gttcagaggc gccctctgaa aaacagaaca gatttcctgg ttcctctgct
     3901 gagctctgtc ttaacagtgg cttgggtctg ttgcttggtg acagccttct actggtgtgt
     3961 gcggaagcgg cggcggaagc ccagcagcca cacccactct gcccctgagg acaacaccac
     4021 caacaatgtg cgcgaacagc tgaaccaaat caaaaacccc atcgagaaac acggagccaa
     4081 cacagtcccc atcaaggatt atgagaacaa aaactccaaa atgtcaaaaa tcaggacaca
     4141 caactcggaa gtggaagagg atgatatgga taagcaccag cagaaagtcc ggtttgccaa
     4201 acagcccgtg tacacgctgg tagacagaga ggagaaggcc cccagcggca cgccgacaaa
     4261 acacccgaac tggacaaaca aacaggacaa cagagacttg gaaagtgccc agagcttgaa
     4321 ccggatggag tacatcgtag aattctgcag atatccagca cagtggcggc cgctcgagga
     4381 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     4441 caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga
     4501 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     4561 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     4621 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     4681 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     4741 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     4801 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     4861 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     4921 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4981 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     5041 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac
     5101 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     5161 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5221 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5281 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     5341 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     5401 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5461 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5521 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5581 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5641 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5701 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     5761 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5821 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5881 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5941 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     6001 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6061 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     6121 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     6181 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     6241 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     6301 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     6361 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     6421 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     6481 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     6541 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6601 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6661 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6721 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6781 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6841 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6901 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6961 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     7021 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     7081 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     7141 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     7201 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     7261 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     7321 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     7381 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     7441 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     7501 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7561 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7621 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7681 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7741 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7801 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7861 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7921 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     7981 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8041 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8101 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     8161 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     8221 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8281 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     8341 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8401 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     8461 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8521 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8581 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8641 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8701 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8761 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8821 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8881 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8941 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     9001 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     9061 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     9121 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     9181 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     9241 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     9301 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     9361 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     9421 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9481 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     9541 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     9601 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     9661 tgaccgacaa ttgcatgaag aatctgctta gg
//
