LOCUS       V046587                 8787 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046587
VERSION     V046587
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..3467
                     /label="Aftph(NM_181411)"
                     /note="Aftph(NM_181411)"
                     /gene="Aftph"
     CDS             3474..3539
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3563..4151
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4217..4265
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4467..4895
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4909..5238
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5305..6099
                     /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    6275..6396
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6445..6461)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6469..6485
                     /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(6493..6523)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6538..6559
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6847..7435)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7606..8466)
                     /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(8467..8571)
                     /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 ccaccatgga gccagacatc attcgaatgt
      781 actcttcctc ccccccacct ctagacaacg gagcagagga tgatgaggag gatgagtttg
      841 gggagtttgg tgggttttcg gaagtcagcc cttctggtgt agggtttgtt gattttgaca
      901 caccagatta tactcgtccc aaggaagact ttgtaccttc aaaccacttt atgccaattc
      961 atgaatattc agaagatgta gatagcctta caagctttaa gtctgttcag aatggtaatg
     1021 ataaggatat cactgctgaa ctttctactc ctgtgaaaag tcaatctgat gttgtacttt
     1081 ctaccaccag caaagaaatg attccatcta aaactttgga tccttccatc gatggtatgg
     1141 aaagcctgga agacttagat aaagtagtgg tgcagggacc aagcactggg caactcagaa
     1201 gtttctctcc tggagatttt agaactgata agaacattgt tcaccaaacc aagcagttag
     1261 agagctgcaa cggtgaaaag cctccttgtc tggaaattct aacaaacggg tttgcaggtt
     1321 tggaaactgt aaatcctcag ggaacagatg acctggacaa tgtggcagat tcaaagggat
     1381 cgaagcctct taacacttgt ggcaccgagt gcattttaga gtctgcagct agtcatgcta
     1441 cggaatttgc agatttttct acattttccc aaacagaaag gacacaactg gaagaaatag
     1501 aatgtccagt tttaaatgat ggtgatacac tgaccattca gggaaacagc aaaggcccca
     1561 gagtcaagga actaaactgt gtgaaagaag tgactttgga tggaagcttt gaggatacgg
     1621 ggaacactga aagagaacat caggtctgcg tctcagaaat ccatgccgta gctgacagag
     1681 ggctcagtgt tgaaaaacaa gaccttcaaa cactgcaaca ggatgagttt ctaaattcaa
     1741 ggattcagtc tgaggcttgg agcttggtag actcaagtga aaattcagaa gccattacaa
     1801 aggagcggtg taaaatggaa aaaaatgact tatttgcctc taagtgtgct gacttatcca
     1861 tggattctgt taaaacttca gacgttaatg aaattggttc ttccaaagaa gaaaacagaa
     1921 agctgactaa tcccaaaagc ccagaccctg atcccacagg acaaaatgct ttggatgatt
     1981 ctgctgcaag catgaaaaat ggtgacagtg ggaatggctt tgtgacttgt catgatacca
     2041 atgaggatga ttttggtgac tttggtacag ccaatggcac cactccaccc tttgttacta
     2101 gtactcaaga ttcaatgagt gacgtcactt ttgaagactc ctcagagcat tttctacatc
     2161 ttagtgaacc aggtgatgat tttggagaat ttgaggatac aaatgctgtt tcttgtcaag
     2221 aggaaatgag atttactgag tcagacctaa gacagacttc tgatggctta tcagaggaat
     2281 gtccattggc aggagagtct ggtggaaaag acagtaaacc tgactcaaaa ctgaaaaatg
     2341 gtcaagacag tgaatttgga gattttgatt ctgtgccaaa tactcaaggt agtgcttttc
     2401 aagactcgga tgattttgca gacttcagtt cagcaggtcc tagccaagct gtagattgga
     2461 atgcttttga ggatgaacag aaagatggtt gttcttgggc tgcctttggg gatcagcagg
     2521 agactgagtc tcatcatcta aaggaagtct ggcagtcaca gaggacagat gaaactatgg
     2581 gcactctggg aacccctaag atgcacagtg tttcttcagc agcttccaaa ggagcagttg
     2641 ctagtggcca tttgcaggaa ccaggcactt cagttcagac agctttactc aaccggttgg
     2701 agagaatttt cgaagcctgt tttccgtctg tgtttgtccc tgatgtggaa gaggaggttt
     2761 cttccctgaa gcacttgctg gagacacaca gctcgccagc aaaaacaaga gaggccttag
     2821 ctgaccgtgg tgagttgagg ggtgtatgga ccgaactgca ggacatccat gatgcacatg
     2881 gtttgcggta ccagtggggt ggctcacata gcaacaagaa gcttttgtgc tccttgggaa
     2941 tagacactcg aaacattcta ttcactggca ataagaagca acctgttata gtgcccatgt
     3001 atgcagcagg attgggaatg ttagagccca ccaaggaacc attaaaacca ctatctgctg
     3061 cagaaaaaat tgcttctatt ggtcagacaa ccgttatgac accagaaata aacacgtgta
     3121 catctgatcc attccaggag tctctcccac ctgtccagtt tgactggagt agcagtggcc
     3181 ttactaaccc tttagatggt gtggatccag agttgtatga gttaacaact gctaagctgg
     3241 agacctccac ctcaagcctc agagtgactg atgcatttgc gaagctcatg tctacagtgg
     3301 aaaagacaag cacgtcaacc aggaaaccaa aaagagagga gcacctgagt gaggaagcca
     3361 tgaaggtgat cgccagcctt ccagacctga ccttcatgca tgccaaggtg ttgatgttcc
     3421 cagccactct gacaccttcc atgagctccc aagaacaagc agacgcggga tccgactaca
     3481 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     3541 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     3601 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3661 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3721 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3781 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3841 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3901 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3961 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4021 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4081 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4141 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     4201 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     4261 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4321 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4381 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     4441 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4501 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4561 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4621 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4681 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4741 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4801 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     4861 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4921 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4981 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5041 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5101 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5161 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5221 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     5281 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5341 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5401 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5461 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5521 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5581 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5641 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5701 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5761 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5821 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5881 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5941 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6001 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6061 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     6121 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6181 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6241 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6361 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6421 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6481 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6541 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6601 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6661 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6721 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6781 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7021 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7261 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7501 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7561 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7621 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7681 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7741 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7801 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7861 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7921 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7981 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8041 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8101 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8161 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8221 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8281 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8341 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8401 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8461 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8521 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8581 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     8641 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     8701 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     8761 gacaattgca tgaagaatct gcttagg
//
