LOCUS       V024559                 7545 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024559
VERSION     V024559
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             705..2201
                     /label="CLK2(NM_001294338)"
                     /note="CLK2(NM_001294338)"
                     /gene="CLK2"
     CDS             2208..2297
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2321..2909
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2975..3023
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3225..3653
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3667..3996
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4063..4857
                     /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    5033..5154
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5203..5219)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5227..5243
                     /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(5251..5281)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5296..5317
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5605..6193)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6364..7224)
                     /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(7225..7329)
                     /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 gtggaatcgc caccatgccg catcctcgaa
      721 ggtaccactc ctcagagcga ggcagccggg ggagttaccg tgaacactat cggagccgaa
      781 agcataagcg acgaagaagt cgctcctggt caagtagtag tgaccggaca cgacggcgtc
      841 ggcgagagga cagctaccat gtccgttctc gaagcagtta tgatgatcgt tcgtccgacc
      901 ggagggtgta tgaccggcga tactgtggca gctacagacg caacgattat agccgggatc
      961 ggggagatgc ctactatgac acagactatc ggcattccta tgaatatcag cgggagaaca
     1021 gcagttaccg cagccagcgc agcagccgga ggaagcacag acggcggagg aggcgcagcc
     1081 ggacatttag ccgctcatct tcgcagcaca gcagccggag agccaagagt gtagaggacg
     1141 acgctgaggg ccacctcatc taccacgtcg gggactggct acaagagcga tatgaaatcg
     1201 ttagcacctt aggagagggg accttcggcc gagttgtaca atgtgttgac catcgcaggg
     1261 gtggggctcg agttgccctg aagatcatta agaatgtgga gaagtacaag gaagcagctc
     1321 gacttgagat caacgtgcta gagaaaatca atgagaaaga ccctgacaac aagaacctct
     1381 gtgtccagat gtttgactgg tttgactacc atggccacat gtgtatctcc tttgagcttc
     1441 tgggccttag caccttcgat ttcctcaaag acaacaacta cctgccctac cccatccacc
     1501 aagtgcgcca catggccttc cagctgtgcc aggctgtcaa gttcctccat gataacaagc
     1561 tgacacatac agacctcaag cctgaaaata ttctgtttgt gaattcagac tatgagctca
     1621 cctacaacct agagaagaag cgagatgagc gcagtgtgaa gagcacagct gtgcgggtgg
     1681 tagactttgg cagtgccacc tttgaccatg agcaccatag caccattgtc tccactcgcc
     1741 attaccgagc accagaagtc atccttgagt tgggctggtc acagccttgt gatgtgtgga
     1801 gtataggctg catcatcttt gaatactatg tgggattcac cctcttccag acccatgaca
     1861 acagagagca tctagccatg atggaaagga tcttgggtcc tatcccttcc cggatgatcc
     1921 gaaagacaag aaagcagaaa tatttttacc ggggtcgcct ggattgggat gagaacacat
     1981 cagctgggcg ctatgttcgt gagaactgca aaccgctgcg gcggtatctg acctcagagg
     2041 cagaggaaca ccaccagctc ttcgatctga ttgaaagcat gctagagtat gaaccagcta
     2101 agcggctgac cttgggtgaa gcccttcagc atcctttctt cgcccgcctt cgggctgagc
     2161 cgcccaacaa gttgtgggac tccagtcggg atatcagtcg gctcgaggaa caaaaactaa
     2221 tatcagagga agatttggaa cagaaactaa tcagtgaaga agatttagag caaaagttaa
     2281 tttccgagga ggacttatag tgagggcccg atatctcgac aatcaacctc tggattacaa
     2341 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2401 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2461 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2521 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     2581 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     2641 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     2701 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     2761 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     2821 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     2881 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     2941 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     3001 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     3061 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     3121 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     3181 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3241 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3301 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3361 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     3421 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3481 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3541 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     3601 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3661 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3721 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3841 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3901 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3961 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4021 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4081 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4141 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4201 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4261 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4321 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4381 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4441 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4501 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4561 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4621 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4681 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4741 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4801 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4861 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4921 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4981 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5041 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5101 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5161 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5221 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5281 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5341 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5401 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5461 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5521 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5581 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5641 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5701 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5761 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     5821 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5881 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5941 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6001 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     6061 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6121 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6181 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6241 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6301 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6361 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6421 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6481 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6541 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6601 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6661 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6721 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6781 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6841 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6901 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6961 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7021 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7081 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7141 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7201 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7261 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7321 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     7381 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     7441 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     7501 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
