LOCUS       V028395                 7897 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028395
VERSION     V028395
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..1032
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     CDS             1039..3417
                     /label="SF3A1(NM_005877)"
                     /note="SF3A1(NM_005877)"
                     /gene="SF3A1"
     polyA_signal    3497..3721
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3767..4195
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4209..4538
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4605..5399
                     /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    5573..5694
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5743..5759)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5767..5783
                     /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(5791..5821)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5836..5857
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6145..6730)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6901..7761)
                     /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(7762..7866)
                     /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 aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt ggtaagccta tccctaaccc tctcctcggt
     1021 ctcgattcta caggtaccat gccggccgga cccgtgcagg cggtgccccc gccgccgccc
     1081 gtgcccacgg agcccaaaca gcccacagaa gaagaagcat cttcaaagga ggattctgca
     1141 ccttctaagc cagttgtggg gattatttac cctcctccag aggtcagaaa tattgttgac
     1201 aagactgcca gctttgtggc cagaaacggg cctgaatttg aagctaggat ccgacagaac
     1261 gagatcaaca accccaagtt caactttctg aaccccaatg acccttacca tgcctactac
     1321 cgccacaagg tcagcgagtt caaggaaggg aaggctcagg agccgtccgc cgccatcccc
     1381 aaggtcatgc agcagcagca gcagaccacc cagcagcagc tgccccagaa ggtccaagcc
     1441 caagtaatcc aagagaccat cgtgcccaaa gagcctcctc ctgagtttga gttcattgct
     1501 gatcctccct ctatctcagc cttcgacttg gatgtggtga agctgacggc tcagtttgtg
     1561 gccaggaatg ggcgccagtt tctgacccag ctgatgcaga aagagcagcg caactaccag
     1621 tttgactttc tccgcccaca gcacagcctc ttcaactact tcacgaagct agtggaacag
     1681 tacaccaaga tcttgattcc acccaaaggt ttattttcaa agctcaagaa agaggctgaa
     1741 aacccccgag aagttttgga tcaggtgtgt taccgagtgg aatgggccaa attccaggaa
     1801 cgtgagagga agaaggaaga agaggagaag gagaaggagc gggtggccta tgctcagatc
     1861 gactggcatg attttgtggt ggtggaaaca gtggacttcc aacccaatga gcaagggaac
     1921 ttccctcccc ccaccacgcc agaggagctg ggggcccgaa tcctcattca ggagcgctat
     1981 gaaaagtttg gggagagtga ggaagttgag atggaggtcg agtctgatga ggaggatgac
     2041 aaacaggaga aggcggagga gcctccttcc cagctggacc aggacaccca agtacaagat
     2101 atggatgagg gttcagatga tgaagaagaa gggcagaaag tgcccccacc cccagagaca
     2161 cccatgcctc cacctctgcc cccaactcca gaccaagtca ttgtccgcaa ggattatgat
     2221 cccaaagcct ccaagccctt gcctccagcc cctgctccag atgagtatct tgtgtccccc
     2281 attactgggg agaagatccc cgccagcaaa atgcaggaac acatgcgcat tggacttctt
     2341 gaccctcgct ggctggagca gcgggatcgc tccatccgtg agaagcagag cgatgatgag
     2401 gtgtacgcac caggtctgga tattgagagc agcttgaagc agttggctga gcggcgtact
     2461 gacatcttcg gtgtagagga aacagccatt ggtaagaaga tcggtgagga ggagatccag
     2521 aagccagagg aaaaggtgac ctgggatggc cactcaggca gcatggcccg gacccagcag
     2581 gctgcccagg ccaacatcac cctccaggag cagattgagg ccattcacaa ggccaaaggc
     2641 ctggtgccag aggatgacac taaagagaag attggcccca gcaagcccaa tgaaatccct
     2701 caacagccac cgccaccatc ttcagccacc aacatcccca gctcggctcc acccatcact
     2761 tcagtgcccc gaccacccac aatgccacct ccagttcgta ctacagttgt ctccgcagta
     2821 cccgtcatgc cccggccccc aatggcatct gtggtccggc tgcccccagg ctcagtgatc
     2881 gcccccatgc cgcccatcat ccacgcgccc agaatcaacg tggtgcccat gcctccctcg
     2941 gcccctccta ttatggcccc ccgcccaccc cccatgattg tgccaacagc ctttgtgcct
     3001 gctccacctg tggcacctgt cccagctcca gccccaatgc cccctgtgca tcccccacct
     3061 cccatggaag atgagcccac ctccaaaaaa ctgaagacag aggacagcct catgccagag
     3121 gaggagttcc tgcgcagaaa caagggtcca gtgtccatca aagtccaggt gcccaacatg
     3181 caggataaga cggaatggaa actgaatggg caggtgctgg tcttcaccct cccactcacg
     3241 gaccaggtct ctgtcattaa ggtgaagatt catgaagcca caggcatgcc tgcagggaaa
     3301 cagaagctac agtatgaggg tatcttcatc aaagattcca actcactggc ttactacaac
     3361 atggccaatg gcgcagtcat ccacctggcc ctcaaggaga gaggcgggag gaagaagtag
     3421 gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     3481 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3541 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3601 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3661 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3721 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3781 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3841 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3901 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3961 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4021 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4081 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4141 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4201 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4261 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4321 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4381 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4441 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4501 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4561 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4621 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4681 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4741 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4801 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4861 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4921 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4981 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5041 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5101 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5161 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5221 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5281 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5341 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5401 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5461 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5521 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5581 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5641 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5701 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5761 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5821 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5881 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5941 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     6001 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     6061 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6121 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     6181 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     6241 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     6301 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     6361 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6421 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6481 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6541 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6601 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6661 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6721 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6781 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6841 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6901 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6961 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7021 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7081 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7141 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7201 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7261 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7321 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7381 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7441 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7501 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7561 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7621 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7681 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7741 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7801 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7861 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
