LOCUS       V024675                 6563 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024675
VERSION     V024675
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..2053
                     /label="CRLF2(NM_022148)"
                     /note="CRLF2(NM_022148)"
                     /gene="CRLF2"
     CDS             2060..2125
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2163..2387
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2433..2861
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2875..3204
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3271..4065
                     /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    4239..4360
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4409..4425)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4433..4449
                     /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(4457..4487)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4502..4523
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4811..5396)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5567..6427)
                     /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(6428..6532)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggggcggc tggttctgct
      961 gtggggagct gccgtctttc tgctgggagg ctggatggct ttggggcaag gaggagcagc
     1021 agaaggagta cagattcaga tcatctactt caatttagaa accgtgcagg tgacatggaa
     1081 tgccagcaaa tactccagga ccaacctgac tttccactac agattcaacg gtgatgaggc
     1141 ctatgaccag tgcaccaact accttctcca ggaaggtcac acttcggggt gcctcctaga
     1201 cgcagagcag cgagacgaca ttctctattt ctccatcagg aatgggacgc accccgtttt
     1261 caccgcaagt cgctggatgg tttattacct gaaacccagt tccccgaagc acgtgagatt
     1321 ttcgtggcat caggatgcag tgacggtgac gtgttctgac ctgtcctacg gggatctcct
     1381 ctatgaggtt cagtaccgga gccccttcga caccgagtgg cagtccaaac aggaaaatac
     1441 ctgcaacgtc accatagaag gcttggatgc cgagaagtgt tactctttct gggtcagggt
     1501 gaaggctatg gaggatgtat atgggccaga cacataccca agcgactggt cagaggtgac
     1561 atgctggcag agaggcgaga ttcgggatgc ctgtgcagag acaccaacgc ctcccaaacc
     1621 aaagctgtcc aaatttattt taatttccag cctggccatc cttctgatgg tgtctctcct
     1681 ccttctgtct ttatggaaat tatggagagt gaagaagttt ctcattccca gcgtgccaga
     1741 cccgaaatcc atcttccccg ggctctttga gatacaccaa gggaacttcc aggagtggat
     1801 cacagacacc cagaacgtgg cccacctcca caagatggca ggtgcagagc aagaaagtgg
     1861 ccccgaggag cccctggtag tccagttggc caagactgaa gccgagtctc ccaggatgct
     1921 ggacccacag accgaggaga aagaggcctc tgggggatcc ctccagcttc cccaccagcc
     1981 cctccaaggc ggtgatgtgg tcacaatcgg gggcttcacc tttgtgatga atgaccgctc
     2041 ctacgtggcg ttgctcgagg actacaaaga ccatgacggt gattataaag atcatgacat
     2101 cgactacaag gatgacgatg acaagtagtg agggcccgtt taaacccgct gatcagcctc
     2161 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2221 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2281 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2341 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2401 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2461 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2521 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2581 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2641 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2701 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2761 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2821 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2881 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2941 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3001 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3061 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3121 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3181 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3241 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3301 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3361 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3421 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3481 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3541 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3601 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3661 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3721 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3781 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3841 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3901 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3961 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4021 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4081 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4141 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4201 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4261 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4321 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4381 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4441 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4501 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4561 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4621 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4681 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4741 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4801 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4861 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4921 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4981 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5041 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5101 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5161 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5221 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5281 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5341 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5401 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5461 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5521 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5581 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5641 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5701 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5761 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5821 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5881 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5941 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6001 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6061 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6121 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6181 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6241 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6301 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6361 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6421 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6481 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6541 cccgaaaagt gccacctgac gtc
//
