LOCUS       V019121                 7355 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019121
VERSION     V019121
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..2035
                     /label="RPS6KB1(NM_001272043)"
                     /note="RPS6KB1(NM_001272043)"
                     /gene="RPS6KB1"
     CDS             2042..2107
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2131..2719
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2785..2833
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3035..3463
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3477..3806
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3873..4667
                     /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    4843..4964
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5013..5029)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5037..5053
                     /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(5061..5091)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5106..5127
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5415..6003)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6174..7034)
                     /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(7035..7139)
                     /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 ccatgaggcg acgaaggagg cgggacggct tttacccagc
      721 cccggacttc cgagacaggg aagctgagga catggcagga gtgtttgaca tagacctgga
      781 ccagccagag gacgcgggct ctgaggatga gctggaggag gggggtcagt taaatgaaag
      841 catggaccat gggggagttg gaccatatga acttggcatg gaacattgtg agaaatttga
      901 aatctcagaa actagtgtga acagagggcc agaaaaaatc agaccagaat gttttgagct
      961 acttcgggta cttggtcaag ggggctatgg aaaggttttt caagtacgaa aagtaacagg
     1021 agcaaatact gggaaaatat ttgccatgaa ggtgcttaaa aaggcaatga tagtaagaaa
     1081 tgctaaagat acagctcata caaaagcaga acggaatatt ctggaggaag taaagcatcc
     1141 cttcatcgtg gatttaattt atgcctttca gactggtgga aaactctacc tcatccttga
     1201 gtatctcagt ggaggagaac tatttatgca gttagaaaga gagggaatat ttatggaaga
     1261 cactgcctgc ttttacttgg cagaaatctc catggctttg gggcatttac atcaaaaggg
     1321 gatcatctac agagacctga agccggagaa tatcatgctt aatcaccaag gtcatgtgaa
     1381 actaacagac tttggactat gcaaagaatc tattcatgat ggaacagtca cacacacatt
     1441 ttgtggaaca atagaataca tggcccctga aatcttgatg agaagtggcc acaatcgtgc
     1501 tgtggattgg tggagtttgg gagcattaat gtatgacatg ctgactggag cacccccatt
     1561 cactggggag aatagaaaga aaacaattga caaaatcctc aaatgtaaac tcaatttgcc
     1621 tccctacctc acacaagaag ccagagatct gcttaaaaag ctgctgaaaa gaaatgctgc
     1681 ttctcgtctg ggagctggtc ctggggacgc tggagaagtt caagctcatc cattctttag
     1741 acacattaac tgggaagaac ttctggctcg aaaggtggag ccccccttta aacctctgtt
     1801 gcaatctgaa gaggatgtaa gtcagtttga ttccaagttt acacgtcaga cacctgtcga
     1861 cagcccagat gactcaactc tcagtgaaag tgccaatcag gtctttctgg gttttacata
     1921 tgtggctcca tctgtacttg aaagtgtgaa agaaaagttt tcctttgaac caaaaatccg
     1981 atcacctcga agatttattg gcagcccacg aacacctgtc agtactgcta tgtgcctcga
     2041 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     2101 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     2161 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2221 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2281 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2341 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2401 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2461 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2521 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2581 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2641 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2701 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     2761 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     2821 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     2881 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     2941 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     3001 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3061 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3121 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3181 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3241 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3301 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3361 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     3421 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3481 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3541 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3601 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3661 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3721 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3781 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     3841 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3901 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3961 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4021 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4081 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4141 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4201 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4261 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4321 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4381 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4441 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4501 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4561 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4621 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4681 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4741 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4801 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     4861 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4921 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     4981 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5041 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5101 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5161 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5221 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5281 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5341 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5401 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5461 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5521 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5581 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     5641 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5701 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5761 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5821 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5881 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5941 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6001 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6061 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6121 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6181 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6241 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6301 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6361 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6421 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6481 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6541 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6601 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6661 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6721 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6781 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6841 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6901 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6961 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7021 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7081 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7141 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     7201 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     7261 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     7321 gcttgaccga caattgcatg aagaatctgc ttagg
//
