LOCUS       V023702                 7771 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023702
VERSION     V023702
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      925..934
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             934..975
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     CDS             982..3261
                     /label="TFRC(NM_003234)"
                     /note="TFRC(NM_003234)"
                     /gene="TFRC"
     CDS             3262..3291
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     polyA_signal    3338..3562
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3608..4036
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4050..4379
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4446..5240
                     /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    5414..5535
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5584..5600)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5608..5624
                     /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(5632..5662)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5677..5698
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5986..6571)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6742..7602)
                     /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(7603..7707)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgggtaagc ctatccctaa ccctctcctc
      961 ggtctcgatt ctacagatat catgatggat caagctagat cagcattctc taacttgttt
     1021 ggtggagaac cattgtcata tacccggttc agcctggctc ggcaagtaga tggcgataac
     1081 agtcatgtgg agatgaaact tgctgtagat gaagaagaaa atgctgacaa taacacaaag
     1141 gccaatgtca caaaaccaaa aaggtgtagt ggaagtatct gctatgggac tattgctgtg
     1201 atcgtctttt tcttgattgg atttatgatt ggctacttgg gctattgtaa aggggtagaa
     1261 ccaaaaactg agtgtgagag actggcagga accgagtctc cagtgaggga ggagccagga
     1321 gaggacttcc ctgcagcacg tcgcttatat tgggatgacc tgaagagaaa gttgtcggag
     1381 aaactggaca gcacagactt caccggcacc atcaagctgc tgaatgaaaa ttcatatgtc
     1441 cctcgtgagg ctggatctca aaaagatgaa aatcttgcgt tgtatgttga aaatcaattt
     1501 cgtgaattta aactcagcaa agtctggcgt gatcaacatt ttgttaagat tcaggtcaaa
     1561 gacagcgctc aaaactcggt gatcatagtt gataagaacg gtagacttgt ttacctggtg
     1621 gagaatcctg ggggttatgt ggcgtatagt aaggctgcaa cagttactgg taaactggtc
     1681 catgctaatt ttggtactaa aaaagatttt gaggatttat acactcctgt gaatggatct
     1741 atagtgattg tcagagcagg gaaaatcacc tttgcagaaa aggttgcaaa tgctgaaagc
     1801 ttaaatgcaa ttggtgtgtt gatatacatg gaccagacta aatttcccat tgttaacgca
     1861 gaactttcat tctttggaca tgctcatctg gggacaggtg acccttacac acctggattc
     1921 ccttccttca atcacactca gtttccacca tctcggtcat caggattgcc taatatacct
     1981 gtccagacaa tctccagagc tgctgcagaa aagctgtttg ggaatatgga aggagactgt
     2041 ccctctgact ggaaaacaga ctctacatgt aggatggtaa cctcagaaag caagaatgtg
     2101 aagctcactg tgagcaatgt gctgaaagag ataaaaattc ttaacatctt tggagttatt
     2161 aaaggctttg tagaaccaga tcactatgtt gtagttgggg cccagagaga tgcatggggc
     2221 cctggagctg caaaatccgg tgtaggcaca gctctcctat tgaaacttgc ccagatgttc
     2281 tcagatatgg tcttaaaaga tgggtttcag cccagcagaa gcattatctt tgccagttgg
     2341 agtgctggag actttggatc ggttggtgcc actgaatggc tagagggata cctttcgtcc
     2401 ctgcatttaa aggctttcac ttatattaat ctggataaag cggttcttgg taccagcaac
     2461 ttcaaggttt ctgccagccc actgttgtat acgcttattg agaaaacaat gcaaaatgtg
     2521 aagcatccgg ttactgggca atttctatat caggacagca actgggccag caaagttgag
     2581 aaactcactt tagacaatgc tgctttccct ttccttgcat attctggaat cccagcagtt
     2641 tctttctgtt tttgcgagga cacagattat ccttatttgg gtaccaccat ggacacctat
     2701 aaggaactga ttgagaggat tcctgagttg aacaaagtgg cacgagcagc tgcagaggtc
     2761 gctggtcagt tcgtgattaa actaacccat gatgttgaat tgaacctgga ctatgagagg
     2821 tacaacagcc aactgctttc atttgtgagg gatctgaacc aatacagagc agacataaag
     2881 gaaatgggcc tgagtttaca gtggctgtat tctgctcgtg gagacttctt ccgtgctact
     2941 tccagactaa caacagattt cgggaatgct gagaaaacag acagatttgt catgaagaaa
     3001 ctcaatgatc gtgtcatgag agtggagtat cacttcctct ctccctacgt atctccaaaa
     3061 gagtctcctt tccgacatgt cttctggggc tccggctctc acacgctgcc agctttactg
     3121 gagaacttga aactgcgtaa acaaaataac ggtgctttta atgaaacgct gttcagaaac
     3181 cagttggctc tagctacttg gactattcag ggagctgcaa atgccctctc tggtgacgtt
     3241 tgggacattg acaatgagtt tgaacaaaaa ctaatatcag aggaagattt gtagctcgag
     3301 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3361 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3421 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3481 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3541 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3601 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3661 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3721 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3781 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3841 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3901 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3961 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4021 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4081 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4141 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4201 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4261 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4321 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4381 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4441 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4501 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4561 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4621 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4681 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4741 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4801 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4861 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4921 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4981 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5041 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5101 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5161 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5221 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5281 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5341 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5401 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5461 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5521 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5581 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5641 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5701 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5761 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5821 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5881 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5941 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6001 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6061 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6121 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6181 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6241 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6301 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6361 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6421 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6481 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     6541 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6601 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6661 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6721 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6781 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6841 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6901 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6961 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7021 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7081 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7141 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7201 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7261 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7321 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7381 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7441 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7501 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7561 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7621 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7681 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7741 cggatcggga gatctcccga tcccctatgg t
//
