LOCUS       V023354                 8256 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023354
VERSION     V023354
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..2936
                     /label="Gpd2(NM_012736)"
                     /note="Gpd2(NM_012736)"
                     /gene="Gpd2"
     CDS             2943..3008
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3032..3620
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3686..3734
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3936..4364
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4378..4707
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4774..5568
                     /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    5744..5865
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5914..5930)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5938..5954
                     /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(5962..5992)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6007..6028
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6316..6904)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7075..7935)
                     /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(7936..8040)
                     /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 ctcggatccg ccaccatggc gtttcaaaag gtagtgaaag
      781 ggactattct tatgggtgga ggagctctgg ccactgtttt gggactctct cagtttgctc
      841 actacagaag gaagcaagta agcctggcgt atgtggaagc agcaacatgc ttctcagagc
      901 ccgtgaacag ggagcctcct tccagagaag ctcagctcat gactctgcag aacacatcgg
      961 aatttgacat cctcgtcatt ggaggaggag ccacaggatg tggctgtgcg ctagacgccg
     1021 tcaccagagg actgaaaaca gcccttgtag agagaaatga cttcgcatca gggactagca
     1081 gcagaagcac taaattgatc catggtggcg tacgatacct ccagaaggct atcacgaact
     1141 tggatgttga acagtatagg atggtgaaag aggcccttca tgaacgtgcc aacttactag
     1201 aaatcgctcc ccatctatca gctccgctgc ctataatgct gccactttat aagtggtggc
     1261 agttacctta ttactgggtg ggaatcaaga tgtatgatct ggttgcagga agtcattgcc
     1321 tgaagagcag ttatgtcctc agtaaatcca gagcccttga gcatttcccc atgctccaga
     1381 aggacaagct ggtcggagcc attgtctact atgatggaca gcacaatgac gcccggatga
     1441 acctcgccat cgccctcacc gctgccaggt acggggctgc cacggccaat tacatggagg
     1501 tggtgagctt gctcaagaag acagaccccg aaactgggaa agagcgcgtg agtggtgccc
     1561 ggtgcaagga cgtgctcaca gggcatgagt ttaacgtgag agctaaatgc gttatcaatg
     1621 ccaccggccc tttcacagac tccgtgcgca aaatggatga taacgatgtt gttcccatct
     1681 gccagcccag tgcaggggtt cacattgtga tgcccgggta ctacagcccc gagaacatgg
     1741 gacttcttga tcctgcaacc agtgatggca gagttatttt cttcttgcct tgggagaaga
     1801 tgacaattgc gggtaccact gattcgccaa cggacgtcac gcaccatcct attccttcgg
     1861 aagacgacat taacttcatc ttaaatgaag tgcggaacta cctgagttgc gacgttgaag
     1921 tgagaagagg ggatgtcctg gcagcgtgga gtggcattcg cccgcttgtt accgatccca
     1981 agtctgcaaa cactcagtcc atctctcgaa atcatgttgt ggaagtcagc gacagtggcc
     2041 tcatcacaat agcaggtggg aagtggacca catatcgctc catggcagaa gatactgtga
     2101 acaaagctgt caagcttcac aacttgaatg caggacccag taggaccgtt gggctcttcc
     2161 ttcaaggagg caaagattgg agccccacac tctacatcag gcttgtccag gactatgggc
     2221 ttgagagtga ggttgcacaa cacctggcca aaacctatgg tgacaaggct ttcgacgtgg
     2281 ccaaaatggc aagtgtgact ggaaaacggt ggcctgttgt tggagtgcgt cttgtgtcag
     2341 aatttccgta catcgaagca gaggtgaaat atgggattaa ggagtacgca tgcaccgctg
     2401 ttgacatgat ttcacgtcgc acccgcctgg cctttcttaa tgttcaggct gcagaggaag
     2461 ccctgcccaa gattgttgag ttaatgggaa gagaattgaa ctggagtgaa ttgaggaaac
     2521 aggaagaact tgaaacagcc acgaggtttc tgtactatga aatgggctat aagtctcgaa
     2581 cagaacaact cacagatagt actgaaatca gcctgctgcc tccagacatc gatcggtaca
     2641 agaagagatt tcacatgttt gatgaagacg aaaaaggctt cattaccatt gttgatgttc
     2701 aacgtgtgct agagagtatc aatgtacaaa tggacgaaga cacactacat gaaattctct
     2761 gtgaagtaga cttgaacaag aatggtcagg ttgagctcca tgagtttctg cagctgatga
     2821 gcgcagttca tacaggaagg gtgtctggaa gccggcttgc catcctcatg aagactgccg
     2881 aggagaactt ggaccgtaga gttccaatcc ccgtggatcg tagttgtgga ggattgctcg
     2941 aggactacaa agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg
     3001 atgacaagta gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga
     3061 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3121 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3181 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3241 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3301 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3361 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3421 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     3481 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3541 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3601 cctttgggcc gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca
     3661 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt
     3721 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     3781 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     3841 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     3901 agatctgcgc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3961 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4021 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4081 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4141 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4201 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4261 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     4321 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4381 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4441 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4501 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4561 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4621 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4681 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4741 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4801 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4861 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4921 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4981 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5041 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5101 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5161 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5221 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5281 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5341 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5401 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5461 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5521 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5581 ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5641 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5701 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5761 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5821 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5881 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5941 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6001 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6061 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6121 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6181 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6241 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6301 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6361 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6421 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6481 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     6541 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6601 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6661 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6721 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6781 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6841 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6901 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6961 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7021 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7081 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7141 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7201 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7261 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7321 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7381 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7441 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7501 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7561 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7621 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7681 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7741 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7801 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7861 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7921 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7981 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8041 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     8101 tggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     8161 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     8221 ggcttgaccg acaattgcat gaagaatctg cttagg
//
