LOCUS       V028889                 7954 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028889
VERSION     V028889
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             706..2610
                     /label="SYK(NM_001174167)"
                     /note="SYK(NM_001174167)"
                     /gene="SYK"
     CDS             2617..2706
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2730..3318
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3384..3432
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3634..4062
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4076..4405
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4472..5266
                     /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    5442..5563
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5612..5628)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5636..5652
                     /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(5660..5690)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5705..5726
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6014..6602)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6773..7633)
                     /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(7634..7738)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggc cagcagcggc
      721 atggctgaca gcgccaacca cctgcccttc tttttcggca acatcacccg ggaggaggca
      781 gaagattacc tggtccaggg gggcatgagt gatgggcttt atttgctgcg ccagagccgc
      841 aactacctgg gtggcttcgc cctgtccgtg gcccacggga ggaaggcaca ccactacacc
      901 atcgagcggg agctgaatgg cacctacgcc atcgccggtg gcaggaccca tgccagcccc
      961 gccgacctct gccactacca ctcccaggag tctgatggcc tggtctgcct cctcaagaag
     1021 cccttcaacc ggccccaagg ggtgcagccc aagactgggc cctttgagga tttgaaggaa
     1081 aacctcatca gggaatatgt gaagcagaca tggaacctgc agggtcaggc tctggagcag
     1141 gccatcatca gtcagaagcc tcagctggag aagctgatcg ctaccacagc ccatgaaaaa
     1201 atgccttggt tccatggaaa aatctctcgg gaagaatctg agcaaattgt cctgatagga
     1261 tcaaagacaa atggaaagtt cctgatccga gccagagaca acaacggctc ctacgccctg
     1321 tgcctgctgc acgaagggaa ggtgctgcac tatcgcatcg acaaagacaa gacagggaag
     1381 ctctccatcc ccgagggaaa gaagttcgac acgctctggc agctagtcga gcattattct
     1441 tataaagcag atggtttgtt aagagttctt actgtcccat gtcaaaaaat cggcacacag
     1501 ggaaatgtta attttggagg ccgtccacaa cttccaggtt cccatcctgc gacttggtca
     1561 gcgggtggaa taatctcaag aatcaaatca tactccttcc caaagcctgg ccacagaaag
     1621 tcctcccctg cccaagggaa ccggcaagag agtactgtgt cattcaatcc gtatgagcca
     1681 gaacttgcac cctgggctgc agacaaaggc ccccagagag aagccctacc catggacaca
     1741 gaggtgtacg agagccccta cgcggacccc gaggagatca ggcccaagga ggtttacctg
     1801 gaccgaaagc tgctgacgct ggaagacaaa gaactgggct ctggtaattt tggaactgtg
     1861 aaaaagggct actaccaaat gaaaaaagtt gtgaaaaccg tggctgtgaa aatactgaaa
     1921 aacgaggcca atgaccccgc tcttaaagat gagttattag cagaagcaaa tgtcatgcag
     1981 cagctggaca acccgtacat cgtgcggatg atcgggatat gcgaggccga gtcctggatg
     2041 ctggttatgg agatggcaga acttggtccc ctcaataagt atttgcagca gaacagacat
     2101 gtcaaggata agaacatcat agaactggtt catcaggttt ccatgggcat gaagtacttg
     2161 gaggagagca attttgtgca cagagatctg gctgcaagaa atgtgttgct agttacccaa
     2221 cattacgcca agatcagtga tttcggactt tccaaagcac tgcgtgctga tgaaaactac
     2281 tacaaggccc agacccatgg aaagtggcct gtcaagtggt acgctccgga atgcatcaac
     2341 tactacaagt tctccagcaa aagcgatgtc tggagctttg gagtgttgat gtgggaagca
     2401 ttctcctatg ggcagaagcc atatcgaggg atgaaaggaa gtgaagtcac cgctatgtta
     2461 gagaaaggag agcggatggg gtgccctgca gggtgtccaa gagagatgta cgatctcatg
     2521 aatctgtgct ggacatacga tgtggaaaac aggcccggat tcgcagcagt ggaactgcgg
     2581 ctgcgcaatt actactatga cgtggtgaac ctcgaggaac aaaaactaat atcagaggaa
     2641 gatttggaac agaaactaat cagtgaagaa gatttagagc aaaagttaat ttccgaggag
     2701 gacttatagt gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa
     2761 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2821 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2881 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2941 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     3001 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     3061 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     3121 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     3181 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     3241 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     3301 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat
     3361 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     3421 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3481 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     3541 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     3601 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3661 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3721 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3781 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3841 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3901 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3961 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     4021 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     4081 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4141 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4201 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4261 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4321 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     4381 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     4441 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4501 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4561 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4621 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4681 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4741 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4801 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4861 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4921 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4981 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5041 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5101 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5161 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5221 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5281 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5341 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5401 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5461 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5521 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     5581 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5641 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5701 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5761 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5821 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5881 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5941 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6001 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6061 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6121 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6181 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     6241 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6301 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6361 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6421 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6481 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6541 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6601 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6661 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6721 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6781 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6841 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6901 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6961 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7021 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7081 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7141 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7201 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7261 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7321 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7381 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7441 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7501 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7561 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7621 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7681 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7741 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     7801 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     7861 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     7921 cttgaccgac aattgcatga agaatctgct tagg
//
