LOCUS       V021710                 7359 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021710
VERSION     V021710
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"
     regulatory      655..664
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             664..729
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             733..2070
                     /label="Klf5(NM_009769)"
                     /note="Klf5(NM_009769)"
                     /gene="Klf5"
     misc_feature    2135..2723
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2789..2837
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3039..3467
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3481..3810
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3877..4671
                     /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    4847..4968
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5017..5033)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5041..5057
                     /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(5065..5095)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5110..5131
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5419..6007)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6178..7038)
                     /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(7039..7143)
                     /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 ttaagccacc
      661 atggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
      721 gatgacaagc ttatgcccac gcgggtgctg accatgagcg cccgcctggg accactgccc
      781 cagccgccgg ccgcgcagga cgagcccgtg ttcgcgcagc tcaagccggt gctgggcgct
      841 gcgaacccgg cccgcgacgc ggcgctcttc tccggagacg atctgaaaca cgcgcaccac
      901 cacccgcctg cgccgccgcc agccgctggc ccgcgactgc cctcggagga gctggtccag
      961 acaagatgtg aaatggagaa gtatctgacc cctcagctcc ctccagttcc gataatttca
     1021 gagcataaaa agtatagacg agacagtgcc tcagtggtag accagttctt cactgacact
     1081 gaaggcatac cttacagcat caacatgaac gtcttcctcc ctgacatcac tcacctgaga
     1141 actggcctct acaaatccca gagaccatgc gtaacacaga tcaagacaga acctgttacc
     1201 attttcagcc accagagcga gtcgacggcc cctcctcctc ctccggcccc cacccaggct
     1261 ctccccgagt tcactagtat cttcagctcc caccagacca cagcgccacc acaggaggtg
     1321 aacaatatct tcatcaaaca agaacttcct ataccagatc ttcatctctc tgtcccttcc
     1381 cagcagggcc acctgtacca gctgttgaat acaccggatc tagacatgcc cagttcgaca
     1441 aaccagacgg cagtaatgga cacccttaat gtttctatgg caggccttaa cccacacccc
     1501 tctgctgttc cacagacgtc aatgaaacag ttccagggca tgcccccttg cacgtacacc
     1561 atgccaagtc agtttcttcc acagcaggcc acctactttc ccccgtcacc accaagctca
     1621 gagcctggaa gtcccgatag acaagctgag atgctgcaga atctcacccc acctccgtcc
     1681 tatgccgcta caattgcttc caaactggcg attcacaacc caaatttacc tgccactctg
     1741 ccagttaatt cgccaactct cccacctgtc agatacaaca gaaggagtaa cccggatctg
     1801 gagaagcgac gtatccactt ctgcgattat aatggttgca caaaagttta tacaaagtcg
     1861 tctcacttaa aagctcacct gaggactcat acgggcgaga agccctacaa gtgcacctgg
     1921 gagggctgcg actggaggtt tgcccggtcg gatgagctga cccgccacta caggaagcac
     1981 acgggcgcca agccgttcca gtgcatggtg tgccaacgca gcttctcccg ctccgaccac
     2041 ctcgcgctgc acatgaagcg ccaccagaac tagaattctg cagatatcca gcacagtggc
     2101 ggccgctcga gtctagaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     2161 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2221 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2281 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2341 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2401 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2461 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2521 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     2581 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2641 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2701 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     2761 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     2821 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     2881 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     2941 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     3001 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3061 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3121 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3181 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3241 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3301 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3361 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     3421 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3481 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3541 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3601 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3661 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3721 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3781 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3841 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3901 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3961 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4021 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4081 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4141 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4201 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4261 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4321 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4381 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4441 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4501 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4561 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4621 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4681 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4741 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4801 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4861 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4921 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4981 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5041 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5101 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5161 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5221 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5281 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5341 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5401 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5461 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5521 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5581 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     5641 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5701 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5761 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5821 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5881 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5941 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6001 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6061 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6121 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6181 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6241 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6301 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6361 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6421 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6481 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6541 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6601 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6661 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6721 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6781 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6841 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6901 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6961 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7021 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7081 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7141 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     7201 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7261 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     7321 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
