LOCUS       V042383                 9487 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042383
VERSION     V042383
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             964..4977
                     /label="PFAS(NM_012393)"
                     /note="PFAS(NM_012393)"
                     /gene="PFAS"
     CDS             4984..5049
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5087..5311
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5357..5785
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5799..6128
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6195..6989
                     /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    7163..7284
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7333..7349)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7357..7373
                     /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(7381..7411)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7426..7447
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7735..8320)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8491..9351)
                     /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(9352..9456)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatgtccc cagtccttca cttctatgtt cgtccctctg gccatgaggg ggcagcccct
     1021 ggacacactc ggaggaaact gcaagggaaa ctgccagagc tgcagggcgt cgagactgaa
     1081 ctgtgctaca acgtgaactg gacagctgag gccctcccca gtgctgagga gacaaagaag
     1141 ctgatgtggc tgtttggttg ccccttactg ctggatgatg ttgctcggga gtcctggctc
     1201 cttcctggct ccaatgacct gctgctggag gtcgggccca ggctgaactt ctccacccca
     1261 acatccacca acatcgtgtc agtgtgccgc gccactgggc tggggcctgt ggatcgtgtg
     1321 gagaccaccc ggcgctaccg gctctcgttt gcccaccccc cgtcagctga ggtggaagcc
     1381 attgctctgg ctaccctgca cgaccggatg acagagcagc acttccccca tcccatccag
     1441 agtttctccc ctgagagcat gccggaaccc ctcaatggcc ctatcaatat actgggtgag
     1501 ggccggcttg cgctggagaa ggccaaccag gagcttggtc tggctttaga ctcttgggac
     1561 ctagacttct acaccaagcg cttccaggag ctacagcgga acccgagcac tgtggaggcc
     1621 tttgacttgg cgcagtccaa tagcgagcac agccgacact ggttcttcaa gggccagctc
     1681 cacgtggatg ggcagaagct ggtgcactca ctgtttgagt ccatcatgag cacccaggaa
     1741 tcctcgaacc ccaacaacgt cctcaaattc tgtgataaca gcagtgcaat ccagggaaag
     1801 gaagtccgat tcctacggcc tgaggacccc acacggccaa gccgcttcca gcaacagcaa
     1861 gggctgagac atgttgtctt cacagcagag actcacaact ttcccacagg agtatgcccc
     1921 tttagtggtg caaccactgg cacagggggc cggattcgag atgtccagtg cacaggccgc
     1981 ggggcccacg tggtggctgg cactgccggc tattgctttg gaaatctgca tattccaggt
     2041 tacaatctgc cctgggagga tccaagcttc cagtatcctg ggaattttgc ccggcccctg
     2101 gaggttgcca ttgaagccag taatggagct tctgactatg gcaacaagtt tggggaacca
     2161 gtgctggctg gcttcgcccg ctccttgggc ctccagctcc cagacggcca gcggcgtgag
     2221 tggatcaagc ccatcatgtt tagtgggggc attgggtcca tggaagctga ccacataagc
     2281 aaggaggccc cagagccagg catggaagtt gtaaaggttg gaggtcccgt ctacaggatt
     2341 ggagttggag gtggagctgc ttcatctgtg caggtgcagg gagataacac cagtgacctg
     2401 gactttgggg ctgtgcagcg aggagacccg gagatggaac agaagatgaa ccgtgtgatc
     2461 agggcttgtg tggaggcccc caagggaaac cccatctgca gccttcatga tcagggcgct
     2521 ggtggcaatg gcaatgtcct aaaagagctg agtgacccag ctggagccat catttacacc
     2581 agccgcttcc agcttgggga cccaaccctg aatgccctgg aaatctgggg ggctgagtac
     2641 caggaatcaa atgctcttct gctgaggtcc cccaaccggg acttcctgac tcatgtcagt
     2701 gcccgtgaac gttgcccggc ttgcttcgtg ggcaccatca ctggagaccg gagaatagtg
     2761 ctggtggacg atcgggagtg tcctgtcaga agaaatggcc agggggatgc ccccccgaca
     2821 cccctgccaa cccctgtgga cctggagctc gaatgggtgc tgggcaagat gcctcggaag
     2881 gagttcttcc tgcagaggaa gccccccatg ctgcagcctc tggccttgcc cccagggctg
     2941 agcgtgcacc aggctctgga gagggttctg aggctgcccg ccgtggccag caagcgctac
     3001 ctcaccaata aggtggaccg ctctgtggga ggcctggtgg cccagcagca gtgcgtgggg
     3061 cccctgcaaa ctcctctggc agatgtagcg gttgtggcac tgagccatga ggagctcata
     3121 ggggctgcca cagccttggg agaacagcca gtcaagagcc tgctggaccc aaaagtcgcc
     3181 gcccggctgg ccgtggccga agccctcacc aacctggtgt ttgctctggt cactgacctc
     3241 cgggatgtga agtgtagcgg gaactggatg tgggcagcca agctcccagg ggagggcgca
     3301 gctttggcgg atgcctgtga ggctatggtg gcagtgatgg cagccctggg tgtggcagtg
     3361 gatggtggca aggactccct cagcatggct gctcgggttg gcactgagac cgtgcgggct
     3421 cctgggtcac tggtcatctc agcctatgcc gtctgcccag acatcacagc cactgtgacc
     3481 ccagacctca agcatcctga agggagaggc catctgctct atgtggctct gagccctggg
     3541 cagcaccggc tcgggggcac agctctggcc cagtgcttct cccagcttgg ggaacaccct
     3601 ccagacctgg accttcctga gaacttggtg cgggccttca gcatcactca ggggctgctg
     3661 aaagaccgcc tcctctgctc aggccacgat gtcagtgacg gaggcctcgt cacatgcctg
     3721 ctggagatgg cctttgctgg aaattgcggg ctacaggtgg atgtgcctgt ccccagggtt
     3781 gatgtcctgt ctgtgctgtt cgctgaggag ccaggcctcg tgctggaggt gcaggagcca
     3841 gacctggccc aggtgctgaa gcgttaccgg gatgctggcc tccattgcct ggagctgggc
     3901 cacacaggcg aggccgggcc ccacgccatg gtccgggtgt cagtgaacgg ggctgtggtt
     3961 ctggaggagc ctgttgggga gctgcgagcc ctctgggagg agacgagttt ccagctggac
     4021 cggctacagg cagagcctcg ctgtgtggca gaggaggaac ggggcctgag ggagcggatg
     4081 gggcccagct attgcctgcc ccccaccttt cccaaagcct ccgtgccccg tgagcctggt
     4141 ggtcccagcc cccgagtcgc catcttgcga gaggagggca gtaatggaga ccgggagatg
     4201 gccgatgcct tccacttagc tgggtttgag gtatgggacg tgaccatgca ggacctctgc
     4261 tctggggcaa ttgggctgga cactttccgt ggcgtggcct tcgtgggcgg cttcagctat
     4321 gcagatgtcc tgggctctgc caaagggtgg gcagctgctg tgacctttca tcccagggct
     4381 ggggctgagc tgaggcgctt ccggaagcgg ccagacacct tcagcctggg cgtgtgtaat
     4441 ggctgtcaac tgctggctct gctcggctgg gtgggaggcg accccaatga ggatgctgca
     4501 gagatgggcc ctgactccca gccagcccgg ccaggccttc tgctacgcca caacctgtct
     4561 gggcgctacg agtctcgctg ggccagcgtg cgtgtggggc ctgggccagc cctgatgctg
     4621 cgagggatgg agggcgccgt gctgcccgtg tggagtgcgc acggggaagg ttacgtagca
     4681 ttttcttctc cggaactcca agctcagatt gaggccaggg gcttggctcc actgcactgg
     4741 gctgatgatg acgggaaccc cacagagcag taccctctga atcccaatgg gtccccaggg
     4801 ggcgtggctg gcatctgctc ctgtgatggc cgccacctgg ctgtcatgcc tcaccctgag
     4861 cgggccgtta ggccttggca gtgggcatgg cgaccccctc catttgatac tctgaccacc
     4921 tccccctggc tccagctctt tatcaatgcc cgaaactgga ccctggaagg gagctgcctc
     4981 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     5041 gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     5101 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     5161 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     5221 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     5281 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     5341 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5401 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5461 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5521 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5581 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5641 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5701 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5761 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     5821 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5881 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5941 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6001 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6061 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6121 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     6181 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     6241 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     6301 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     6361 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     6421 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     6481 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     6541 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     6601 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     6661 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     6721 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     6781 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     6841 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6901 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6961 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     7021 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     7081 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     7141 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     7201 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     7261 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     7321 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7381 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7441 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7501 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7561 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7621 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7681 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7741 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7801 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7861 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7921 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7981 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8041 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8101 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8161 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8221 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     8281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     8341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     8401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     8461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     8521 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     8581 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     8641 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     8701 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     8761 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     8821 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     8881 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     8941 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     9001 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     9061 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     9121 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     9181 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     9241 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     9301 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     9361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     9481 tgacgtc
//
