LOCUS       V042098                 7629 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042098
VERSION     V042098
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2309
                     /label="Ccnl2(NM_207678)"
                     /note="Ccnl2(NM_207678)"
                     /gene="Ccnl2"
     CDS             2316..2381
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2405..2993
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3059..3107
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3309..3737
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3751..4080
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4147..4941
                     /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    5117..5238
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5287..5303)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5311..5327
                     /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(5335..5365)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5380..5401
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5689..6277)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6448..7308)
                     /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(7309..7413)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc ggcggcggcg gccggggctg
      781 cggggttgat ggctccggcg tcggcggcct gctcttccgg ctcggcggga gcggccccgg
      841 ggtctcaggg ggtgctgatc ggcgacaggc tctactcggg cgtgctcatc accttggaaa
      901 actgcctgct gcccgacgac aagctccgct tcacgccgtc catgtcgagc ggcctcgacg
      961 tcgatacgga gacgggactc cgcgtcgtgg gctgcgagct catccaggca gccggcatcc
     1021 tgctccgcct gccgcaggtg gccatggcta cagggcaggt gctgttccag cggttctttt
     1081 acactaagtc cttcgtgaaa cattccatgg agcatgtgtc catggcttgt gttcacctgg
     1141 cttccaaaat agaagaggct ccgagacgaa tcagggatgt catcaatgta ttccatcgcc
     1201 ttcggcatct gagagagaaa aagaagcccg tgcctctggt gcttgatcaa gagtatgtta
     1261 acctaaagaa ccagattata aaggcagaga gacgggttct caaggagttg ggcttttgtg
     1321 tccatgtgaa gcaccctcac aagataatcg ttatgtacct tcaggtgtta gaatgtgagc
     1381 gtaatcaaca cctggtccag actgcatgga actacatgaa tgacagcctt cgcacagatg
     1441 tctttgtccg gttccagcct gaaagcattg cttgtgcctg tatctacctt gctgcccgga
     1501 cactggagat ccctttaccc aatcgtccac attggtttct tttgtttgga gcaactgaag
     1561 aagaaattca agaaatttgc tttaaaatct tgcagcttta tacacggaaa aaggttgatt
     1621 tgacgcatct ggagagtgaa gtggagaaga gaaagcacgc cattgaagaa gcaaaggccc
     1681 gagcaaaggg cctgctccca gggactgcgc caggcctgga cagtgctgcg gggttctcgc
     1741 ctgctcccaa gctggaatcc ccaaaagaag gtaaaggagg caagccttcc ccgccctctg
     1801 gaaagagcgc caaacggaaa atggaaggcc caaagaaagc ccagggccac agccctgtaa
     1861 atggcttgct gaaagggcag gagagtcgaa gtcaaagcag gagtcgtgag cagagctact
     1921 caaggtcccc atcacggtcc gcttctccaa agagaaggaa aagtgatagt ggctctacct
     1981 caggtgggtc caagtcacag agtcgttctc ggagccgaag tgactctcct ccaaggcagg
     2041 tacaccgagg tgctccctac aaaggctcag aagtgagggg ctcccggaaa tcaaaggact
     2101 gcaagtacct cacccagaag ccacacaagt ctcgtagccg gagctcctct cgttcccgaa
     2161 gccggtcacg agagcggacg gacaattctg gaaaatacaa gaagaaaagt cattactata
     2221 gagatcaaag acgtgagcgc tcaaggtcat atgagcgcac aggccatcgc tatgagaggg
     2281 accaccctgg acacagcagg catcggaggg gatccgacta caaagaccat gacggtgatt
     2341 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     2401 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2461 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2521 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2581 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2641 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2701 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2761 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     2821 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2881 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2941 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     3001 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     3061 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     3121 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     3181 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     3241 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     3301 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3361 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3421 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     3481 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3541 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3601 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3661 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     3721 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3841 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3901 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3961 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4021 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4081 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4141 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4201 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4261 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4321 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4381 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4441 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4501 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4561 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4621 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4681 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4741 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4801 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4861 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4921 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     4981 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5041 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5101 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5161 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5221 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5281 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5341 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5401 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5461 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5521 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5581 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5641 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5701 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5761 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5821 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5881 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5941 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6001 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6061 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6121 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6181 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6241 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6301 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6361 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6421 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6481 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6541 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6601 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6661 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6721 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6781 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6841 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6901 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6961 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7021 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7081 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7141 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7201 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7261 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7321 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7381 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7441 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     7501 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7561 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     7621 ctgcttagg
//
